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Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract γ-polyglutamic acid (γ-PGA), as an environmentally sustainable material, is extensive applied in agriculture for enhancing water and fertilizer utilization efficiency, augmenting crop yield, and ameliorating soil conditions. However, the effect of γ-PGA in conjunction with sesame cake fertilizer on the soil environment remains uncertain.The aim of this study is to investigate the effect of γ-PGA on soil nutrients, water use efficiency (WUE) and nitrogen use efficiency (NUE), and maize yield across various levels of sesame cake fertilizer. Additionally, the study seeks to identify the optimal ratio to establish a theoretical and practical foundation for sustainable agricultural development and the promotion of ecological agriculture. Through field experiments, nine treatments were established, comprising three levels of sesame cake fertilizer application rates (B1 = 900 kg/hm2 for low fertility, B2 = 1100 kg/hm2 for medium fertility, and B3 = 1300 kg/hm2 for high fertility) and three levels of γ-PGA application rates (R1 = 200 kg/hm2, R2 = 400 kg/hm2, and R3 = 600 kg/hm2). The results can be outlined as follows: (1) When γ-PGA application rate increased, total nitrogen (TN) exhibited a synergistic effect under B1 treatment, but an antagonistic effect under B2 and B3 treatments. At the 6-leaf stage (V6), 12-leaf stage (V12), and tasseling stage (VT), available phosphorus (AP) exhibited antagonistic effects. However, at the filling stage (R2) and maturity stage (R6), AP in B1 and B2 treatments at various depths underwent partial transformation into a synergistic effect. The levels of available potassium exhibited a notable antagonistic effect, leading to a decrease in harvest index (HI). B2 treatment demonstrated superior results compared to the B1 and B3 treatments, with the highest levels observed under B2R1 treatment; (2) TN content in the 0–40 cm soil layer increased during the filling period, and it was uniformly distributed in the 40–60 cm soil layer. When the soil AP was located in the 0–60 cm soil layer, there was an increase in AP content during the mature period. Following the tasseling period, different treatments exhibited varying patterns of increase in response to the presence of potassium within the 0–60 cm soil layer. Consequently, in cases where the sesame cake fertilizer content is low, the interaction between γ-PGA can compensate for the deficiency of fertilizer, thereby enhancing water and nitrogen utilization efficiency. The optimal fertilization strategy for enhancing soil nutrient distribution, WUE and NUE, and yield is proposed to be the application of 1100 kg/hm2 sesame cake fertilizer and 200 kg/hm2 γ-PGA.

γ-polyglutamic acid (γ-PGA), as an environmentally sustainable material, is extensive applied in agriculture for enhancing water and fertilizer utilization efficiency, augmenting crop yield, and ameliorating soil conditions. However, the effect of γ-PGA in conjunction with sesame cake fertilizer on the soil environment remains uncertain.The aim of this study is to investigate the effect of γ-PGA on soil nutrients, water use efficiency (WUE) and nitrogen use efficiency (NUE), and maize yield across various levels of sesame cake fertilizer. Additionally, the study seeks to identify the optimal ratio to establish a theoretical and practical foundation for sustainable agricultural development and the promotion of ecological agriculture. Through field experiments, nine treatments were established, comprising three levels of sesame cake fertilizer application rates (B1 = 900 kg/hm2 for low fertility, B2 = 1100 kg/hm2 for medium fertility, and B3 = 1300 kg/hm2 for high fertility) and three levels of γ-PGA application rates (R1 = 200 kg/hm2, R2 = 400 kg/hm2, and R3 = 600 kg/hm2). The results can be outlined as follows: (1) When γ-PGA application rate increased, total nitrogen (TN) exhibited a synergistic effect under B1 treatment, but an antagonistic effect under B2 and B3 treatments. At the 6-leaf stage (V6), 12-leaf stage (V12), and tasseling stage (VT), available phosphorus (AP) exhibited antagonistic effects. However, at the filling stage (R2) and maturity stage (R6), AP in B1 and B2 treatments at various depths underwent partial transformation into a synergistic effect. The levels of available potassium exhibited a notable antagonistic effect, leading to a decrease in harvest index (HI). B2 treatment demonstrated superior results compared to the B1 and B3 treatments, with the highest levels observed under B2R1 treatment; (2) TN content in the 0–40 cm soil layer increased during the filling period, and it was uniformly distributed in the 40–60 cm soil layer. When the soil AP was located in the 0–60 cm soil layer, there was an increase in AP content during the mature period. Following the tasseling period, different treatments exhibited varying patterns of increase in response to the presence of potassium within the 0–60 cm soil layer. Consequently, in cases where the sesame cake fertilizer content is low, the interaction between γ-PGA can compensate for the deficiency of fertilizer, thereby enhancing water and nitrogen utilization efficiency. The optimal fertilization strategy for enhancing soil nutrient distribution, WUE and NUE, and yield is proposed to be the application of 1100 kg/hm2 sesame cake fertilizer and 200 kg/hm2 γ-PGA. [ABSTRACT FROM AUTHOR]

Poly-γ-glutamic acid (γ-PGA) is a carboxylic-acid-rich, bio-derived, water-soluble, edible, hydrating, non-immunogenic polymer produced naturally by several microorganisms. Here, we re-emphasise the ability of Bacillus subtilis natto to naturally produce γ-PGA on whole seaweed, as well as for the yields and chemical properties of the material to be affected by the presence of Mn(2+). Hyaluronic acid (HA) is an extracellular glycosaminoglycan which presents a high concentration of carboxylic acid and hydroxyl groups, being key in fulfilling numerous applications. Currently, there are strong environmental (solvent use), social (non-vegan extraction), and economic factors pushing for the biosynthesis of this material through prokaryotic microorganisms, which is not yet scalable or sustainable. Our study aimed to investigate an innovative raw material which can combine both superior hygroscopicity and UV protection to the cosmetic industry. Comparable hydration effect of commercially available γ-PGA to conventional moisturising agents (HA and glycerol) was observed; however, greater hydration capacity was observed from seaweed-derived γ-PGA. Herewith, successful incorporation of seaweed-derived γ-PGA (0.2–2 w/v%) was achieved for several model cream systems with absorbances reported at 300 and 400 nm. All γ-PGA-based creams displayed shear thinning behaviour as the viscosity decreased, following increasing shear rates. Although the use of commercial γ-PGA within creams did not suggest a significant effect in rheological behaviour, this was confirmed to be a result of the similar molecular weight. Seaweed-derived γ-PGA cream systems did not display any negative effect on model HaCaT keratinocytes by means of in vitro MTT analysis. [ABSTRACT FROM AUTHOR]

Plant microbiota composition changes with the environment and host state, suggesting potential for engineering. However, engineering plant microbiomes is promising but currently undeveloped. This study investigated the role of root-associated bacterial microbiomes in poly-γ-glutamic acid (γ-PGA)-induced plant disease resistance. γ-PGA treatment significantly reduced wilt disease caused by Fusarium oxysporum f. sp. chrysanthemi (Foc). Quantitative PCR analysis revealed a 73.2% reduction in Foc abundance in the roots following γ-PGA exposure. However, the disease suppression effect of γ-PGA was notably weakened in sterilized soils or soils treated with bactericide, indicating the essential role of root-associated microbiomes in this process. 16S rRNA gene amplicon sequencing showed that γ-PGA treatments increased the abundance of Proteobacteria, particularly the family Burkholderiaceae, in the roots. Metabolite analysis further indicated that γ-PGA treatment significantly elevated salicylic acid (SA) levels, suggesting that SA played a critical role in the assembly of the root microbiome under γ-PGA treatment. Further experiments confirmed the antagonistic activity and induced systemic resistance (ISR) of Burkholderia sp. against Fusarium wilt. Burkholderia sp. CM72 was found to enhance plant disease resistance through antibiosis and activation of jasmonic acid (JA)-related pathways. In summary, γ-PGA significantly improved plant disease resistance by modulating the SA pathway and promoted the colonization of beneficial microbiota, particularly with Burkholderia sp. [ABSTRACT FROM AUTHOR]

The growing demand for natural products in the cosmetic industry has led to increased interest in natural polysaccharides that offer biocompatibility, biodegradability, and bioactivity. Microorganisms have emerged as a promising source for large-scale production of these polysaccharides under controlled conditions. Among them, the halophilic bacterium Chromohalobacter canadensis 28 stands out as it produces an exopolymer (Cc EP) consisting of exopolysaccharides (EPS) and poly-gamma-glutamic acid (γ-PGA), making it the first halophilic microorganism known for PGA production. This study explores the potential of crude Cc EP and purified γ-PGA for improving the skin barrier, stimulating collagen and hyaluronic acid production, and facilitating wound closure. Through in-vitro experiments using human epidermal keratinocyte cells (HaCaT) and dermal fibroblast cells (PCS-201-012), both the exopolymers were found to exhibit positive effects on cellular proliferation and the expression of genes related to collagen, hyaluronic acid, involucrin, and filaggrin. Furthermore, in-vitro scratch models demonstrated their ability to enhance wound closure. Based on these promising findings, we propose that Cc EP and purified γ-PGA hold potential as active ingredients in cosmeceutical products. Their ability to promote cell proliferation, improve collagen and hyaluronic acid expression, and enhance wound closure makes them valuable assets for skincare applications. [ABSTRACT FROM AUTHOR]

In order to explore the optimal fermentation conditions for the poly-γ-glutamic acid (γ-PGA) production and sensory quality of fermented natto, the comprehensive score of γ-PGA production and sensory evaluation after natto fermentation was used as an index to optimize the natto fermentation process by single factor test, Plackett-Burman test design, steepest climbing test and Box-Behnken response surface method. The optimum fermentation conditions of natto were determined as follows: Soybean soaking pH7.2, fermentation temperature 37.10 ℃, fermentation time 24.20 h, inoculation amount 4.00%, bottling amount 19.90%. Under the optimized conditions, the production of γ-PGA was 3.65 mg/g, the sensory score was 31.67, and the comprehensive score was 10.04. There was no significant difference between the experimental results and the model prediction values, which was stable and reliable.

BACKGROUND: Natto mucus is mainly composed of poly(γ‐glutamic acid) (γ‐PGA), which affects the sensory quality of natto and has some effective functional activities. The soybean metabolites that cause different γ‐PGA contents in different fermented natto are unclear. RESULTS: In this study, we use untargeted metabolomics to analyze the metabolites of high‐production γ‐PGA natto and low‐production γ‐PGA natto and their fermented substrate soybean. A total of 257 main significantly different metabolites with the same trend among the three comparison groups were screened, of which 114 were downregulated and 143 were upregulated. Through the enrichment of metabolic pathways, the metabolic pathways with significant differences were purine metabolism, nucleotide metabolism, fructose and mannose metabolism, anthocyanin biosynthesis, isoflavonoid biosynthesis and the pentose phosphate pathway. CONCLUSION: For 114 downregulated main significantly different metabolites with the same trend among the three comparison groups, Bacillus subtilis (natto) may directly decompose them to synthesize γ‐PGA. Adding downregulated substances before fermentation or cultivating soybean varieties with the goal of high production of such substances has a great effect on the production of γ‐PGA by natto fermentation. The enrichment analysis results showed the main pathways affecting the production of γ‐PGA by Bacillus subtilis (natto) using soybean metabolites, which provides a theoretical basis for the production of γ‐PGA by soybean and promotes the diversification of natto products. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Previously, a novel Corynebacterium glutamicum strain for the de novo biosynthesis of tailored poly-γ-glutamic acid (γ-PGA) has been constructed by our group. The strain was based on the γ-PGA synthetase complex, PgsBCA, which is the only polyprotein complex responsible for γ-PGA synthesis in Bacillus spp. In the present study, PgsBCA was reconstituted and overexpressed in C. glutamicum to further enhance γ-PGA synthesis. First, we confirmed that all the components (PgsB, PgsC, and PgsA) of γ-PGA synthetase derived from B. licheniformis are necessary for γ-PGA synthesis, and γ-PGA was detected only when PgsB, PgsC, and PgsA were expressed in combination in C. glutamicum. Next, the expression level of each pgsB , pgsC , and pgsA was tuned in order to explore the effect of expression of each of the γ-PGA synthetase subunits on γ-PGA production. Results showed that increasing the transcription levels of pgsB or pgsC and maintaining a medium-level transcription level of pgsA led to 35.44% and 76.53% increase in γ-PGA yield (γ-PGA yield-to-biomass), respectively. Notably, the expression level of pgsC had the greatest influence (accounting for 68.24%) on γ-PGA synthesis, followed by pgsB. Next, genes encoding for PgsC from four different sources (Bacillus subtilis , Bacillus anthracis , Bacillus methylotrophicus , and Bacillus amyloliquefacien s) were tested in order to identify the influence of PgsC-encoding orthologues on γ-PGA production, but results showed that in all cases the synthesis of γ-PGA was significantly inhibited. Similarly, we also explored the influence of gene orthologues encoding for PgsB on γ-PGA production, and found that the titer increased to 17.14 ± 0.62 g/L from 8.24 ± 0.10 g/L when PgsB derived from B. methylotrophicus replaced PgsB alone in PgsBCA from B. licheniformis. The resulting strain was chosen for further optimization, and we achieved a γ-PGA titer of 38.26 g/L in a 5 L fermentor by optimizing dissolved oxygen level. Subsequently, by supplementing glucose, γ-PGA titer increased to 50.2 g/L at 48 h. To the best of our knowledge, this study achieved the highest titer for de novo production of γ-PGA from glucose, without addition of L-glutamic acid, resulting in a novel strategy for enhancing γ-PGA production. [ABSTRACT FROM AUTHOR]

Osseous disease accounts for over half of chronic pathologies, but there is a limited supply of autografts, the gold standard; hence, there is a demand for new synthetic biomaterials. Herein, we present the use of a promising, new dairy-derived biomaterial: whey protein isolate (WPI) in the form of hydrogels, modified with the addition of different concentrations of the biotechnologically produced protein-like polymeric substance poly-γ-glutamic acid (γ-PGA) as a potential scaffold for tissue regeneration. Raman spectroscopic analysis demonstrated the successful creation of WPI-γ-PGA hydrogels. A cytotoxicity assessment using preosteoblastic cells demonstrated that the hydrogels were noncytotoxic and supported cell proliferation from day 3 to 14. All γ-PGA-containing scaffold compositions strongly promoted cell attachment and the formation of dense interconnected cell layers. Cell viability was significantly increased on γ-PGA-containing scaffolds on day 14 compared to WPI control scaffolds. Significantly, the cells showed markers of osteogenic differentiation; they synthesised increasing amounts of collagen over time, and cells showed significantly enhanced alkaline phosphatase activity at day 7 and higher levels of calcium for matrix mineralization at days 14 and 21 on the γ-PGA-containing scaffolds. These results demonstrated the potential of WPI-γ-PGA hydrogels as scaffolds for bone regeneration. [ABSTRACT FROM AUTHOR]

Copyright of Science & Technology of Food Industry is the property of Science & Technology of Food Industry Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Poly-gamma-glutamic acid (PGA) is a naturally occurring biopolymer that has been receiving increasing attention in various fields, including the food, medical, and cosmetic industries. This review provides a comprehensive overview of PGA, discussing its biosynthesis, characteristics, and emerging applications. The biosynthesis of PGA is discussed in detail, highlighting the various microorganisms that are capable of producing PGA and the factors that affect its production. The structure and molecular weight of PGA are also described, as well as its solubility and biodegradability. These properties make PGA a highly versatile material that can be utilized in a wide range of applications. The review also provides an in-depth analysis of the emerging applications of PGA. In the food industry, PGA has been used as a natural preservative, a thickener, and a flavor enhancer. In the medical field, PGA has shown promising results as a drug delivery system and a scaffold for tissue engineering. The challenges and future directions for the production and utilization of PGA are also discussed. This review article covers key aspects, including production process optimization, property improvement, and exploration of novel applications for PGA. It serves as a valuable resource for researchers and industry professionals who wish to explore the potential of PGA in diverse applications. The exceptional properties of PGA make it highly appealing, with a broad spectrum of potential uses across multiple industries. [ABSTRACT FROM AUTHOR]

Currently, the industrial production of γ-polyglutamic acid (γ-PGA) was mainly based on microbial fermentation, but the decolorization technology still needs to be improved. Therefore, in this study, the optimal decolorization conditions and efficiencies were investigated. As the result, D4006 was an ideal resin for the decolorization of γ-PGA fermentation broth. The optimal decolorization pH value and temperature were pH 3 and 40 °C, respectively; ultrafiltration differential pressure, temperature and time were 0.2 MPa, 40 °C, and 6 h, respectively. Moreover, the combination of D4006 (3 %) resin on the basis of ultrafiltration had a better decolorization effect on γ-PGA fermentation broth, and the decolorization rate reached 96.8 %. [ABSTRACT FROM AUTHOR]

Poly-γ-glutamic acid (γ-PGA) is a carboxylic-acid-rich, bio-derived, water-soluble, edible, hydrating, non-immunogenic polymer produced naturally by several microorganisms. Here, we re-emphasise the ability of Bacillus subtilis natto to naturally produce γ-PGA on whole seaweed, as well as for the yields and chemical properties of the material to be affected by the presence of Mn(2+). Hyaluronic acid (HA) is an extracellular glycosaminoglycan which presents a high concentration of carboxylic acid and hydroxyl groups, being key in fulfilling numerous applications. Currently, there are strong environmental (solvent use), social (non-vegan extraction), and economic factors pushing for the biosynthesis of this material through prokaryotic microorganisms, which is not yet scalable or sustainable. Our study aimed to investigate an innovative raw material which can combine both superior hygroscopicity and UV protection to the cosmetic industry. Comparable hydration effect of commercially available γ-PGA to conventional moisturising agents (HA and glycerol) was observed; however, greater hydration capacity was observed from seaweed-derived γ-PGA. Herewith, successful incorporation of seaweed-derived γ-PGA (0.2–2 w/v%) was achieved for several model cream systems with absorbances reported at 300 and 400 nm. All γ-PGA-based creams displayed shear thinning behaviour as the viscosity decreased, following increasing shear rates. Although the use of commercial γ-PGA within creams did not suggest a significant effect in rheological behaviour, this was confirmed to be a result of the similar molecular weight. Seaweed-derived γ-PGA cream systems did not display any negative effect on model HaCaT keratinocytes by means of in vitro MTT analysis.

Plant microbiota composition changes with the environment and host state, suggesting potential for engineering. However, engineering plant microbiomes is promising but currently undeveloped. This study investigated the role of root-associated bacterial microbiomes in poly-γ-glutamic acid (γ-PGA)-induced plant disease resistance. γ-PGA treatment significantly reduced wilt disease caused by Fusarium oxysporum f. sp. chrysanthemi (Foc). Quantitative PCR analysis revealed a 73.2% reduction in Foc abundance in the roots following γ-PGA exposure. However, the disease suppression effect of γ-PGA was notably weakened in sterilized soils or soils treated with bactericide, indicating the essential role of root-associated microbiomes in this process. 16S rRNA gene amplicon sequencing showed that γ-PGA treatments increased the abundance of Proteobacteria, particularly the family Burkholderiaceae, in the roots. Metabolite analysis further indicated that γ-PGA treatment significantly elevated salicylic acid (SA) levels, suggesting that SA played a critical role in the assembly of the root microbiome under γ-PGA treatment. Further experiments confirmed the antagonistic activity and induced systemic resistance (ISR) of Burkholderia sp. against Fusarium wilt. Burkholderia sp. CM72 was found to enhance plant disease resistance through antibiosis and activation of jasmonic acid (JA)-related pathways. In summary, γ-PGA significantly improved plant disease resistance by modulating the SA pathway and promoted the colonization of beneficial microbiota, particularly with Burkholderia sp.

Abstract Background The root systems of higher plants play an important role in plant growth and development. In our present study, it was found that poly-γ-glutamic acid (γ-PGA), an environmentally friendly biomacromolecule, significantly improved root development in maize. Results After treatment with γ-PGA for 7 days, the fresh weight of maize roots was significantly increased and the differences between γ-PGA treated group and control group were mainly caused by the number (higher by 71.87% compared to the control) and length of lateral roots. RNAseq and RT-PCR analyses showed that γ-PGA treatment upregulated the expression of genes related to the synthesis of auxins and auxin signal in maize roots. In addition, γ-PGA promoted the accumulation of plant growth-promoting bacteria, such as Azospirillum, Azohydromonas, Ramlibacter, and Sphingobium (Proteobacteria), Streptomyces (Actinobacteria), Parasegetibacter (Bacteroidetes), and Gemmatimonas (Gemmatimonadetes) in rhizosphere soil and the secretion of auxins. The results of this study deepened our understanding of the effects and mechanism of γ-PGA on maize root development, and as well as highlighted the possibility of using γ-PGA to improve crop growth and soil environment. Conclusions γ-PGA promotes early growth and development of maize roots by inducing the secretion and accumulation of auxin in roots and in rhizosphere soil, and increasing the abundance of plant growth promoting bacteria.

Soils are typically subjected to multiple wetting–drying (WD) cycles due to irrigation and seasonal climate cycles, which directly impact soil pore structure and soil aggregate stability. Poly‐γ‐glutamic acid (γ‐PGA) is a polymer used to improve soil water holding capacity and plant growth. However, the impact of γ‐PGA on soil pore structure requires further research, particularly under WD cycles. Therefore, we investigated the different amounts of γ‐PGA on soil structure, including soil aggregate stability, macropore (>100 μm) structure characteristics and the relationship between macropore characteristics (equivalent pore diameter, pore shape factor, soil porosity, fractal dimension (FD), soil connectivity and the percentage of aggregate content with particle size larger than 0.25 mm) and soil aggregate stability by structural equation modelling (SEM) under WD cycles. A sandy soil and a loam soil were studied, and amended with γ‐PGA at three different concentrations: 0 (P0), 0.4% (P4) and 0.8% (P8) (w/w, %). Results showed that γ‐PGA amendment increased the mean weight diameter (MWD) and the percentage of aggregate content with particle size larger than 0.25 mm (R0.25) under WD cycles in both sandy and loam soils, while the MWD between P4 and P8 did not differ significantly. As the number of WD cycles increased, soil porosity (TP) increased due to an increase in pores of 100–500 μm. With γ‐PGA added to soil, large microporosity (>1000 μm) increased in sandy soil, but decreased in loam soil. In addition, 8WD cycles also increased the FD (2.6%–4.2%) and pore connectivity (Con) compared with 4WD. Structural equation modelling (SEM) revealed that soil pore characteristics accounted for 74% and 98% of the variation in sandy and loam soils, respectively. TP, FD, Con and R0.25 directly contributed to MWD, according to the SEM. These findings improve our understanding of pore characteristics and aggregate stability, which are key factors influencing soil quality when amended with γ‐PGA during the seasonal WD period. [ABSTRACT FROM AUTHOR]

Poly--glutamic acid super absorbent polymer (1-PGA SAP) is a novel type of SAP synthesized by 1-PGA. To study the effect of 1-PGA SAP on the growth of winter wheat and the microenvironment of the soil root zone, five levels of 1-PGA SAP addition (0, 40 kg/hm2, 80 kg/hm2, 120 kg/hm2, 160 kg/hm2) and three different irrigation levels (270 mm, 229.5 mm, and 189 mm) were set in the field. The average soil water content of each treatment increased gradually with increasing 1-PGA SAP addition. The number of soil microorganisms and the activities of soil enzymes increased with increasing 1-PGA SAP addition. The wheat yields without 1-PGA SAP addition were 5873.08 kg/hm2 (270 mm), 5081.65 kg/hm2 (229.5 mm) and 4181.41 kg/hm2 (189 mm) in three different irrigation levels, respectively. Wheat yield increased with increasing 1-PGA SAP addition. The yield of winter wheat increased by 9.00% (270 mm), 17.94% (229.5 mm) and 26.31% (189 mm) respectively compared with the control group when the 1-PGA SAP addition was 160kg/hm2. However, the increasing rate of wheat yield decreased when the 1-PGA SAP addition was more than 80 kg/hm2. [ABSTRACT FROM AUTHOR]

(-)-Epigallocatechin gallate (EGCG) has gained considerable attention owing to its beneficial properties. However, its application as a functional food is restricted due to its instability and low bioavailability. In the present study, a food-derived nanoparticle system based on type A gelatin/γ-PGA was developed to preserve and deliver EGCG. The EGCG/gelatin/γ-PGA nanoparticles had a particle size of 155.1 ± 7.3 nm with a zeta potential of −23.9 ± 0.9 mV. Moreover, the EGCG/gelatin/γ-PGA nanoparticles enhanced the long-term storage stability and sustained antioxidant activity of EGCG compared to EGCG/gelatin nanoparticles. The nanoparticles protected EGCG in simulated gastric fluid containing pepsin while releasing it in simulated intestinal fluid. Additionally, the amount of EGCG transported in the Caco-2 monolayers treated with EGCG/gelatin/γ-PGA nanoparticles was three times higher than that of free EGCG, which might be related to the paracellular pathway and endocytosis. These results suggest that EGCG/gelatin/γ-PGA nanoparticles might be an effective delivery vehicle for EGCG, enhancing its potential applications in the functional food field. [ABSTRACT FROM AUTHOR]

Osseous disease accounts for over half of chronic pathologies, but there is a limited supply of autografts, the gold standard; hence, there is a demand for new synthetic biomaterials. Herein, we present the use of a promising, new dairy-derived biomaterial: whey protein isolate (WPI) in the form of hydrogels, modified with the addition of different concentrations of the biotechnologically produced protein-like polymeric substance poly-γ-glutamic acid (γ-PGA) as a potential scaffold for tissue regeneration. Raman spectroscopic analysis demonstrated the successful creation of WPI-γ-PGA hydrogels. A cytotoxicity assessment using preosteoblastic cells demonstrated that the hydrogels were noncytotoxic and supported cell proliferation from day 3 to 14. All γ-PGA-containing scaffold compositions strongly promoted cell attachment and the formation of dense interconnected cell layers. Cell viability was significantly increased on γ-PGA-containing scaffolds on day 14 compared to WPI control scaffolds. Significantly, the cells showed markers of osteogenic differentiation; they synthesised increasing amounts of collagen over time, and cells showed significantly enhanced alkaline phosphatase activity at day 7 and higher levels of calcium for matrix mineralization at days 14 and 21 on the γ-PGA-containing scaffolds. These results demonstrated the potential of WPI-γ-PGA hydrogels as scaffolds for bone regeneration.

Poly-γ-glutamic acid (γ-PGA) is an emerging biopolymer produced by several Bacillus species. To improve γ-PGA synthesis, metabolic engineering of the production host B. subtilis poses great potential and is facilitated by the convenient genetical amenability of the organism. In this study, a 3.7-fold increase in γ-PGA production using a bdhA, alsSD, pta, yvmC, and cypX deletion mutant with blocked by-product synthesis pathways was obtained. A detailed analysis of intracellular metabolites for reference strains and the γ-PGA-producing deletion strain identified the accumulation of pyruvate and acetyl-CoA in deletion mutants, highlighting the citrate synthase activity as an important metabolic engineering target for further metabolic flux optimization towards γ-PGA synthesis. An in-depth analysis of growth and γ-PGA production with on-line measurement techniques revealed significant variations across cultivations with deletion mutants that are likely caused by culture acidification due to pyruvate accumulation. Despite the observed acidification, the by-product deletion mutants outperformed the reference strains independent of the promoter controlling the PGA synthetase expression. The constructed deletion strains exhibit high γ-PGA production in minimal medium with glucose as sole carbon source as well as in modified Medium E reaching γ-PGA concentrations of 0.57 gL-1 and 14.46 gL-1, respectively. The results presented in this work broaden the understanding of the microbial metabolism during γ-PGA production and will be useful to guide future metabolic engineering for improved γ-PGA production.

Copyright of Irrigation & Drainage is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Recently, human hair keratin has been widely studied and applied in clinical fields due to its good histocompatibility, biocompatibility, and biodegradability. However, the regenerated keratin from human hair cannot be electrospun alone because of its low molecular weight. Herein, gamma polyglutamic acid (γ-PGA) was first selected to fabricate smooth and uniform γ-PGA/keratin composite scaffolds with excellent biocompatibility and biodegradability by electrospinning technology and a chemical cross-linking method in this study. The effect of electrospinning parameters on the structure and morphology, the mechanism of chemical cross-linking, biocompatibility in vitro cell culture experiments, and biodegradability in phosphate-buffered saline buffer solution and trypsin solution of the γ-PGA/keratin electrospun nanofibrous scaffolds (ENS) was studied. The results show that the cross-linked γ-PGA/keratin ENSs had excellent water stability and biodegradability. The γ-PGA/keratin ENSs showed better biocompatibility in promoting cell adhesion and cell growth compared with the γ-PGA ENSs. It is expected that γ-PGA/keratin ENSs will be easily and significantly used in tissue engineering to repair or regenerate materials. [ABSTRACT FROM AUTHOR]

Poly-γ-glutamic acid (γ-PGA) is a biopolymer composed of glutamate monomers. This polypeptide is used in the food, biomedical, and biopharmaceutical industries. It is used for wound healing and surgical suturing and applied as a drug carrier and drug delivery agent. Fermentative production of this polyamide is a usual method accomplished by microbial cultures. The E culture medium is common for the biopolymers synthesis by Bacillus licheniformis ATCC 9945a strain because of the high attained production amounts. Nevertheless, in this study, the culture medium E compounds were modified to attain a novel medium for more production of γ-PGA. The next purpose of the research was more production increasing of γ-PGA by using a novel simultaneous pulsed feeding strategy of citrate and glutamate in the modified E medium. Two pulsed feeding times of 24th and 48th hours with proper amounts of citrate and glutamate resulted in enhanced production of γ-PGA to 119.31 g/l, which is the highest amount among previous researches. This polyamide's physical and chemical structures were investigated by SEM and FTIR, respectively. These analyses revealed a uniform nanostructured polyamide with chemical specifications similar to standard samples. [ABSTRACT FROM AUTHOR]

Poly-γ-glutamic acid (γ-PGA) is an environment-friendly polymeric biomaterial with good water absorption and slow-release. Its metabolites are non-toxic and edible, and it has hence been widely used in a wide range of fields including cosmetics, food and medicine. Its application in agriculture is mainly as a soil conditioner to improve the ability of soil to retain water, as well as a fertilizer synergistic agent. Studies in recent years have concluded that γ-PGA has a potential application in many areas in agricultural production. In this paper, we reviewed its application as a soil conditioner to improve water bioavailability. We reviewed previous studies on the effects of γ-PGA on physicochemical and biological properties of soil, fertilizer leaching and adsorption, the ultimate crop growth and development, as well as nutrient absorption and physiological traits of different crops. We also outline the existing research challenges and the prospects of γ-PGA in its application in agricultural production.

Abstract Background Highly contagious respiratory diseases caused by viral infections are a constantly emerging threat, particularly the elderly with the higher risk of developing serious complications. Vaccines are the best strategy for protection against influenza-related diseases. However, the elderly has lower vaccine efficacy than young population and the age-driven decline of the influenza vaccine efficacy remains unresolved. Objectives This study investigates the effect of an adjuvant, poly-γ-glutamic acid and alum (PGA/Alum) on vaccine efficacy in aged mice (18-months) and its mechanism is investigated using ovalbumin as a model antigen and a commercial pandemic H1N1 (pH1N1) flu vaccine. Antigen trafficking, dendritic cell (DC) activation, and the DC-mediated T cell activation were analyzed via in vivo imaging and flow cytometry. Antigen-specific humoral and cellular immune responses were evaluated in sera and splenocytes from the vaccinated mice. Also, we analyzed gene expression profiles of splenocytes from the vaccinated mice via single-cell transcriptome sequencing and evaluated the protective efficacy against pH1N1 virus challenge. Results Aged mice had lower antigen trafficking and DC activation than younger mice (6-weeks), which was ameliorated by PGA/Alum with increased antigen uptake and DC activation leading to improved antigen-specific IFN-γ+CD8+ T lymphocyte frequencies higher in the vaccinated aged mice, to a similar extent as PGA/Alum adjuvanted vaccine-immunized young mice. The results of single-cell transcriptome sequencing display that PGA/Alum also reduced the proportion of age-associated CD8+ T cell subsets and gene levels of inhibitory regulators in CD8+ T cells, which may play a role in the recovery of CD8+ T cell activation. Finally, PGA/Alum adjuvanted pH1N1 vaccine-immunized aged mice were completely protected (100% survival) compared to aged mice immunized with vaccine only (0% survival) after pH1N1 virus challenge, akin to the efficacy of the vaccinated young mice (100% survival). Conclusions PGA/Alum adjuvanted pH1N1 vaccine-immunized aged mice showed a significant increase in vaccine efficacy compared to aged mice administered with vaccine only. The enhanced vaccine efficacy by PGA/Alum is associated with significant increases of activation of DCs and effector CD8+ T cells and a decrease in age-associated CD8+ T cell proportion of splenocytes. Collectively, PGA/Alum adjuvanted flu vaccine may be a promising vaccine candidate for the elderly.

Abstract Background Compared with other abiotic stresses, drought stress causes serious crop yield reductions. Poly-γ-glutamic acid (γ-PGA), as an environmentally friendly biomacromolecule, plays an important role in plant growth and regulation. Results In this project, the effect of exogenous application of γ-PGA on drought tolerance of maize (Zea mays. L) and its mechanism were studied. Drought dramatically inhibited the growth and development of maize, but the exogenous application of γ-PGA significantly increased the dry weight of maize, the contents of ABA, soluble sugar, proline, and chlorophyll, and the photosynthetic rate under severe drought stress. RNA-seq data showed that γ-PGA may enhance drought resistance in maize by affecting the expression of ABA biosynthesis, signal transduction, and photosynthesis-related genes and other stress-responsive genes, which was also confirmed by RT–PCR and promoter motif analysis. In addition, diversity and structure analysis of the rhizosphere soil bacterial community demonstrated that γ-PGA enriched plant growth promoting bacteria such as Actinobacteria, Chloroflexi, Firmicutes, Alphaproteobacteria and Deltaproteobacteria. Moreover, γ-PGA significantly improved root development, urease activity and the ABA contents of maize rhizospheric soil under drought stress. This study emphasized the possibility of using γ-PGA to improve crop drought resistance and the soil environment under drought conditions and revealed its preliminary mechanism. Conclusions Exogenous application of poly-γ-glutamic acid could significantly enhance the drought resistance of maize by improving photosynthesis, and root development and affecting the rhizosphere microbial community.

A novel approach to fabricate porous mullite ceramics with homogeneous pore size and high-strength using green non-toxic and cost-effective poly- γ -glutamic acid (γ -PGA) gelling system was reported for the first time. Effect of γ -PGA addition, additive amount and solid loading on rheological behavior of the slurries, and microstructure and properties of samples were investigated systematically. By optimizing the solid loading of mullite samples, we are able to get the sample with small pores (< 200 µm) dominating (93.3% of the total pores), and compressive strength of the sample reaches up to 26.62 MPa. In addition, the mullite ceramics exhibited high porosity of 75.7% with low thermal conductivity of 0.279 W/(m·K) at room temperature. This study not only provides a green and non-toxic gelling system but also offers porous mullite ceramics with low thermal conductivity and excellent mechanical strength as an energy-saving thermal insulation material. [ABSTRACT FROM AUTHOR]

Cancer nanovaccines have emerged as an indispensable weapon for tumor treatment. However, insufficient immunogenicity and immunosuppression hamper the therapeutic effects of nanovaccines. Here, biodegradable nanovaccines (OMPP) composed of ovalbumin (OVA)-manganese oxide nanoparticles, amphiphilic poly(γ-glutamic acid) (γ-PGA), and ε-polylysine (PL) are constructed to realize enhanced cancer immunotherapy. Interestingly, amphiphilic γ-PGA and PL could serve as both carriers and immunoadjuvants to promote the cytosolic delivery of antigens and enhance the maturation of dendritic cells. Additionally, taking advantage of the photothermal property of OMPP, immunogenic cell death and in situ release of tumor-associated antigens can be triggered under near-infrared light irradiation for personalized tumor treatment. Moreover, OMPP nanovaccines can efficiently alleviate tumor hypoxia and downregulate programmed death-ligand 1 expression to reprogram the immunosuppressive tumor microenvironment. OMPP-mediated therapy has been shown to provoke robust immune responses to suppress B16-OVA melanoma and prevent postsurgical tumor recurrence. This work presents a facile strategy for the fabrication of nanovaccines by integrating carrier and adjuvant while exploring the inherent properties to promote antigen release and modulate immunosuppression, which demonstrates great potential for effective cancer immunotherapy.

Poly-γ-glutamic acid (γ-PGA) is a versatile biopolymer with widespread applications in the food, pharmaceutical, and medical industries. One of the main challenges in expanding γ-PGA industrial applications is the high cost of production. Developing an efficient and low-cost fermentation process such as bacterial cultivation with pulsed feeding can significantly reduce production costs. Thus, initially, a new pulsed-feeding strategy of citrate and glutamate was developed for γ-PGA production enhancement in the fed-batch culture of Bacillus licheniformis ATCC 9945a. Then, the effects of pulse number, feeding amount, feeding times, the addition time of calcium and manganese solutions, the pH of the added citrate solution, and the concentration of feed stock solutions of pulse-feeds on γ-PGA production were investigated. Under optimal conditions: feeding two pulses at 8 and 24 hours of culture, 20 g citrate and glutamate per liter of culture medium per pulse (about 52 mL of each of citrate and glutamate feeding solutions prepared with a concentration of 384 g/L by adding distilled water) about 88 ± 4 g/L of γ-PGA was obtained. It is one of the highest values ever reported for γ-PGA production with Bacillus licheniformis ATCC 9945a, of course with a much simpler process than the other fed-batch processes. [ABSTRACT FROM AUTHOR]

Milk was co-fermented with Bacillus subtilis HA and Lactiplantibacillus plantarum EJ2014 to produce a dairy ingredient enriched with poly-γ-glutamic acid (γ-PGA) and γ-aminobutyric acid (GABA). The first fermentation of milk with B. subtilis HA resulted in a viscous broth with pH 6.56, 0.26% acidity, 1.40 mg/g tyrosine equivalent, and 17.21 U/g protease activity. The viable cell counts of B. subtilis indicated 8.74 log CFU/mL, and the consistency index of the alkaline fermented milk was 1.82 Pa·sn. In addition, 4.65% mucilage was produced with 35.93% γ-PGA content. The milk co-fermented by L. plantarum indicated 1.34% acidity and pH 4.91. The viable bacterial counts of B. subtilis decreased to 4.44 log CFU/mL, whereas those of L. plantarum increased to 9.42 log CFU/mL. Monosodium glutamate (MSG) as a precursor was effectively converted into γ-PGA by B. subtilis, and then residual MSG was completely converted into GABA by L. plantarum with a yield of 26.15 mg/g. Furthermore, the co-fermented milk produced volatiles, including hexanoic acid, 2,3-butanediol, and acetoin, which may be responsible for its aged cheese-like aroma. [ABSTRACT FROM AUTHOR]

Poly-γ-glutamic acid (γ-PGA) is a bio-derived water-soluble, edible, non-immunogenic nylon-like polymer with the biochemical characteristics of a polypeptide. This Bacillus-derived material has great potential for a wide range of applications, from bioremediation to tunable drug delivery systems. In the context of oral care, γ-PGA holds great promise in enamel demineralisation prevention. The salivary protein statherin has previously been shown to protect tooth enamel from acid dissolution and act as a reservoir for free calcium ions within oral cavities. Its superb enamel-binding capacity is attributed to the L-glutamic acid residues of this 5380 Da protein. In this study, γ-PGA was successfully synthesised from Bacillus subtilis natto cultivated on supplemented algae media and standard commercial media. The polymers obtained were tested for their potential to inhibit demineralisation of hydroxyapatite (HAp) when exposed to caries simulating acidic conditions. Formulations presenting 0.1, 0.25, 0.5, 0.75, 1, 2, 3 and 4% (w/v) γ-PGA concentration were assessed to determine the optimal conditions. Our data suggests that both the concentration and the molar mass of the γ-PGA were significant in enamel protection (p = 0.028 and p < 0.01 respectively). Ion Selective Electrode, combined with Fourier Transform Infra-Red studies, were employed to quantify enamel protection capacity of γ-PGA. All concentrations tested showed an inhibitory effect on the dissolution rate of calcium ions from hydroxyapatite, with 1% (wt) and 2% (wt) concentrations being the most effective. The impact of the average molar mass (M) on enamel dissolution was also investigated by employing commercial 66 kDa, 166 kDa, 440 kDa and 520 kDa γ-PGA fractions. All γ-PGA solutions adhered to the surface of HAp with evidence that this remained after 60 min of continuous acidic challenge. Inductively Coupled Plasma analysis showed a significant abundance of calcium ions associated with γ-PGA, which suggests that this material could also act as a responsive calcium delivery system. We have concluded that all γ-PGA samples tested (commercial and algae derived) display enamel protection capacity regardless of their concentration or average molar mass. However, we believe that γ-PGA D/L ratios might affect the binding more than its molar mass. [ABSTRACT FROM AUTHOR]

Abstract Poly-γ-glutamic acid (γ-PGA) and nattokinase (NK) are the main substances produced by Bacillus subtilis natto in solid-state fermentation and have wide application prospects. We found that our strains had higher activity of nattokinase when soybeans were used as substrate to increase the yield of γ-PGA. Commercial production of γ-PGA and nattokinase requires an understanding of the mechanism of co-production. Here, we obtained the maximum γ-PGA yield (358.5 g/kg, w/w) and highest activity of NK during fermentation and analyzed the transcriptome of Bacillus subtilis natto during co-production of γ-PGA and NK. By comparing changes in expression of genes encoding key enzymes and the metabolic pathways associated with the products in genetic engineering, the mechanism of co-production of γ-PGA and nattokinase can be summarized based on RNA-seq analysis. This study firstly provides new insights into the mechanism of co-production of γ-PGA and nattokinase by Bacillus subtilis natto and reveals potential molecular targets to promote the co-production of γ-PGA and nattokinase.

[Display omitted] • γ-PGA-producing Bacillus subtilis Tamang strain with 43.2% GC content was isolated from kinema. • γ-PGA showed amide groups of glutamic acid, high molecular weight, α-helical conformation. • γ-PGA showed good iron solubilization, ferritin retention in Caco-2 cells. • γ-PGA synthase complex (capBCA) for biosynthesis of γ-PGA detected in genome. • Multiple potential genes were identified without any AMR genes or virulence factors. Kinema , a traditional fermented soybean food from the Himalayas, is well-liked for its sticky texture and flavourful umami taste. Among 175 bacterial strains from spontaneously fermented kinema samples, Bacillus subtilis Tamang strain stood out for its high stickiness and viscosity. The strain's Poly-γ-glutamic acid (γ-PGA) contains various groups of glutamic acid and has a molecular weight of 660 kDa. It demonstrates the ability to solubilize iron, preserve ferritin in Caco-2 cells, and exhibit antibacterial properties. The genome of B. subtilis Tamang is devoid of plasmid elements but does feature nine insert elements. Noteworthy is the presence of unique secondary metabolites with potential antimicrobial effects, such as amyloliquecidin GF610, bogorol A, and thermoactinoamide A. A total of 132 carbohydrate-active enzymes (CAZy) were identified, hinting at possible prebiotic characteristics. The genome analysis revealed genes responsible for γ-PGA production via the capBCA complex. Furthermore, genes associated with fibrinolytic activity, taste enhancement, biopeptides, immunomodulators, and vitamins like B12 and K2 were found, along with probiotics and various health benefits. The genetic material for L-asparaginase production, known for its anti-cancer properties, was also detected, as well as CRISPR-Cas systems. The absence of virulence factors and antimicrobial resistance genes confirms the safety of consuming B. subtilis Tamang as a food-grade bacterium. [ABSTRACT FROM AUTHOR]

Drought stress is the main factor restricting maize yield. Poly-γ-glutamic acid (γ-PGA), as a water-retaining agent and fertilizer synergist, could significantly improve the drought resistance and yield of many crops. However, its high production costs and unclear long-term impact on soil ecology limit its large-scale application. In this study, an environmentally friendly green material γ-PGA was heterologous synthesized in maize for the first time using the synthetic biology method. The genes (PgsA , PgsB , PgsC) participated in γ-PGA synthesis were cloned from Bacillus licheniformis and transformed into maize to produce γ-PGA for the first time. Under drought stress, transgenic maize significantly increased the ear length, ear weight and grain weight by 50 % compared to the control, whereas the yield characteristic of ear weight, grain number per ear, grain weight per ear and 100-grain weight increased by 1.67 %–2.33 %, 3.78 %–13.06 %, 8.41 %–22.06 %, 6.03 %–19.28 %, and 11.85 %–18.36 %, respectively under normal growth conditions. γ-PGA was mainly expressed in the mesophyll cells of maize leaf rosette structure and improved drought resistance and yield by protecting and increasing the expression of genes for the photosynthetic and carbon fixation. This study is an important exploration for maize drought stress molecular breeding and building resource-saving agriculture. [ABSTRACT FROM AUTHOR]

(-)-Epigallocatechin gallate (EGCG) has gained considerable attention owing to its beneficial properties. However, its application as a functional food is restricted due to its instability and low bioavailability. In the present study, a food-derived nanoparticle system based on type A gelatin/γ-PGA was developed to preserve and deliver EGCG. The EGCG/gelatin/γ-PGA nanoparticles had a particle size of 155.1 ± 7.3 nm with a zeta potential of −23.9 ± 0.9 mV. Moreover, the EGCG/gelatin/γ-PGA nanoparticles enhanced the long-term storage stability and sustained antioxidant activity of EGCG compared to EGCG/gelatin nanoparticles. The nanoparticles protected EGCG in simulated gastric fluid containing pepsin while releasing it in simulated intestinal fluid. Additionally, the amount of EGCG transported in the Caco-2 monolayers treated with EGCG/gelatin/γ-PGA nanoparticles was three times higher than that of free EGCG, which might be related to the paracellular pathway and endocytosis. These results suggest that EGCG/gelatin/γ-PGA nanoparticles might be an effective delivery vehicle for EGCG, enhancing its potential applications in the functional food field.

Background: Highly contagious respiratory diseases caused by viral infections are a constantly emerging threat, particularly the elderly with the higher risk of developing serious complications. Vaccines are the best strategy for protection against influenza-related diseases. However, the elderly has lower vaccine efficacy than young population and the age-driven decline of the influenza vaccine efficacy remains unresolved. Objectives: This study investigates the effect of an adjuvant, poly-γ-glutamic acid and alum (PGA/Alum) on vaccine efficacy in aged mice (18-months) and its mechanism is investigated using ovalbumin as a model antigen and a commercial pandemic H1N1 (pH1N1) flu vaccine. Antigen trafficking, dendritic cell (DC) activation, and the DC-mediated T cell activation were analyzed via in vivo imaging and flow cytometry. Antigen-specific humoral and cellular immune responses were evaluated in sera and splenocytes from the vaccinated mice. Also, we analyzed gene expression profiles of splenocytes from the vaccinated mice via single-cell transcriptome sequencing and evaluated the protective efficacy against pH1N1 virus challenge. Results: Aged mice had lower antigen trafficking and DC activation than younger mice (6-weeks), which was ameliorated by PGA/Alum with increased antigen uptake and DC activation leading to improved antigen-specific IFN-γ+CD8+ T lymphocyte frequencies higher in the vaccinated aged mice, to a similar extent as PGA/Alum adjuvanted vaccine-immunized young mice. The results of single-cell transcriptome sequencing display that PGA/Alum also reduced the proportion of age-associated CD8+ T cell subsets and gene levels of inhibitory regulators in CD8+ T cells, which may play a role in the recovery of CD8+ T cell activation. Finally, PGA/Alum adjuvanted pH1N1 vaccine-immunized aged mice were completely protected (100% survival) compared to aged mice immunized with vaccine only (0% survival) after pH1N1 virus challenge, akin to the efficacy of the vaccinated young mice (100% survival). Conclusions: PGA/Alum adjuvanted pH1N1 vaccine-immunized aged mice showed a significant increase in vaccine efficacy compared to aged mice administered with vaccine only. The enhanced vaccine efficacy by PGA/Alum is associated with significant increases of activation of DCs and effector CD8+ T cells and a decrease in age-associated CD8+ T cell proportion of splenocytes. Collectively, PGA/Alum adjuvanted flu vaccine may be a promising vaccine candidate for the elderly. [ABSTRACT FROM AUTHOR]

Poly-γ-glutamic acid (γ-PGA) is an important biopolymer with many applications due to its biodegradable and non-toxic characteristics. γ-PGA is produced industrially by fermentation of Bacillus species. The optimal pH range for producing γ-PGA by Bacillus subtilis PG-001 was firstly studied by glucose fed-batch fermentation with non-controlled pH. Result showed that both cell growth and γ-PGA synthesis were repressed when pH was lower than pH 6. Further investigation with γ-PGA fed-batch fermentation showed that pH 6.5 is more suitable for γ-PGA fermentation than pH 7. Under comparable consumption of glutamic acid and glucose, 11.8 g/L γ-PGA and 0.7 g/g yield were achieved by fermentation at pH 6.5, which was significantly higher than 10.5 g/L and 0.56 g/g yield of fermentation at pH 7. In addition, γ-PGA degradation during later phase of fermentation was repressed at pH 6.5 as 9238cP of final broth viscosity was achieved from fermentation at pH 6.5 while it was only 346 cP for fermentation at pH 7. Finally, a glucose feedback control pH-stat strategy was performed for reducing alkali consumption during γ-PGA fermentation, which further increased final γ-PGA concentration to 15.5 g/L with much higher viscosity (11458 cP); meanwhile the consumption of alkali decreased 57%. The fed-batch γ-PGA fermentation with glucose feedback control pH-stat strategy showed high feasibility for industrial scaling-up. [ABSTRACT FROM AUTHOR]

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Biostimulants can enhance horticultural crop production. However, their application in hydroponically grown lettuce is still limited, and information regarding their relative efficacy is lacking. A greenhouse trial was conducted to address this issue. Five nutrient solution treatments were evaluated on two lettuce cultivars: butterhead and red oak-leaf. The treatments included a half-strength modified Hoagland solution (Hs-H); a full-strength modified Hoagland solution (Fs-H); and Hs-H supplemented with 50 mg L−1 fulvic acid (FA), 334 mg L−1 seaweed extract (SE), or 5 mL L−1 gamma polyglutamic acid (PGA). The results indicated that the shoot biomass observed after biostimulant supplementation was significantly greater than or comparable to that observed with Fs-H. Nutrient solutions supplemented with SE and PGA led to a greater increase in the root biomass than that realized with Hs-H and Fs-H treatments. The Hs-H + FA treatment resulted in the lowest root-to-shoot ratio on a fresh weight basis among all treatments. The nitrate concentration in the shoot was significantly reduced following biostimulant supplementation compared to that realized with Fs-H and Hs-H treatments. Nutrient solutions supplemented with SE and PGA also decreased soluble sugar concentrations compared to that achieved using Hs-H and Fs-H treatments. FA and SE improved nutrient uptake for both cultivars, but PGA had a minimal effect on nutrient uptake. The two cultivars varied in their responses to biostimulant supplementation with regard to biomass, quality traits, and nutrient uptake. This study supports using fulvic acid and seaweed extract, rather than γ-PGA, in hydroponic lettuce production systems. [ABSTRACT FROM AUTHOR]

Lignocellulosic biomass is an important raw material which has been extensively studied for biotechnological applications, but research on its use for the production of γ-polyglutamic acid (γ-PGA) is rarely reported. The present study aimed to improve the release of xylose and glucose from corncob for γ-PGA production using the strain Bacillus subtilis SCP010-1, which could metabolize xylose. The effectiveness of the three chemical reagents NaOH, NH3 and H2O2 pretreatments was investigated by the response surface methodology (RSM). The results showed that maximum glucose and xylose yields of 11.8 ± 0.3 and 17.36 ± 0.6 g/L, respectively, as determined by phloroglucinol-acid and enzymatic methods, were obtained when using 0.986% of NaOH for 2.07 h. Then, the pretreatment solution was subject to further optimize conditions of enzymatic hydrolysis, and the content of glucose and xylose in the corncob hydrolysate reached respectively 15.3 ± 0.3 and 22.34 ± 0.17 g/L under synergism of hemicellulase and cellulase. Finally, the fermentation conditions for γ-PGA production by B. subtilis SCP010-1 were also optimized, and after adding supplemental nutrients to the corncob hydrolysate, a maximum γ-PGA production of 30.035 ± 0.49 g/L was achieved by fed batch fermentation with feeding corncob hydrolysate. Considering the cost of the materials used in this study, nearly 85% of reduction in cost could be achieved, hence indicating that this production plan offers good prospects for industrial production of γ-PGA. More importantly, this process allows the use of environmental-friendly biological resources. [ABSTRACT FROM AUTHOR]

Lignocellulosic biomass is an important raw material which has been extensively studied for biotechnological applications, but research on its use for the production of γ-polyglutamic acid (γ-PGA) is rarely reported. The present study aimed to improve the release of xylose and glucose from corncob for γ-PGA production using the strain Bacillus subtilis SCP010–1, which could metabolize xylose. The effectiveness of the three chemical reagents NaOH, NH 3 and H 2 O 2 pretreatments was investigated by the response surface methodology (RSM). The results showed that maximum glucose and xylose yields of 11.8 ± 0.3 and 17.36 ± 0.6 g/L, respectively, as determined by phloroglucinol-acid and enzymatic methods, were obtained when using 0.986% of NaOH for 2.07 h. Then, the pretreatment solution was subject to further optimize conditions of enzymatic hydrolysis, and the content of glucose and xylose in the corncob hydrolysate reached respectively 15.3 ± 0.3 and 22.34 ± 0.17 g/L under synergism of hemicellulase and cellulase. Finally, the fermentation conditions for γ-PGA production by B. subtilis SCP010–1 were also optimized, and after adding supplemental nutrients to the corncob hydrolysate, a maximum γ-PGA production of 30.035 ± 0.49 g/L was achieved by fed batch fermentation with feeding corncob hydrolysate. Considering the cost of the materials used in this study, nearly 85% of reduction in cost could be achieved, hence indicating that this production plan offers good prospects for industrial production of γ-PGA. More importantly, this process allows the use of environmental-friendly biological resources. [ABSTRACT FROM AUTHOR]

Background: Compared with other abiotic stresses, drought stress causes serious crop yield reductions. Poly-γ-glutamic acid (γ-PGA), as an environmentally friendly biomacromolecule, plays an important role in plant growth and regulation. Results: In this project, the effect of exogenous application of γ-PGA on drought tolerance of maize (Zea mays. L) and its mechanism were studied. Drought dramatically inhibited the growth and development of maize, but the exogenous application of γ-PGA significantly increased the dry weight of maize, the contents of ABA, soluble sugar, proline, and chlorophyll, and the photosynthetic rate under severe drought stress. RNA-seq data showed that γ-PGA may enhance drought resistance in maize by affecting the expression of ABA biosynthesis, signal transduction, and photosynthesis-related genes and other stress-responsive genes, which was also confirmed by RT–PCR and promoter motif analysis. In addition, diversity and structure analysis of the rhizosphere soil bacterial community demonstrated that γ-PGA enriched plant growth promoting bacteria such as Actinobacteria, Chloroflexi, Firmicutes, Alphaproteobacteria and Deltaproteobacteria. Moreover, γ-PGA significantly improved root development, urease activity and the ABA contents of maize rhizospheric soil under drought stress. This study emphasized the possibility of using γ-PGA to improve crop drought resistance and the soil environment under drought conditions and revealed its preliminary mechanism. Conclusions: Exogenous application of poly-γ-glutamic acid could significantly enhance the drought resistance of maize by improving photosynthesis, and root development and affecting the rhizosphere microbial community. [ABSTRACT FROM AUTHOR]

Recently, human hair keratin has been widely studied and applied in clinical fields due to its good histocompatibility, biocompatibility, and biodegradability. However, the regenerated keratin from human hair cannot be electrospun alone because of its low molecular weight. Herein, gamma polyglutamic acid (γ-PGA) was first selected to fabricate smooth and uniform γ-PGA/keratin composite scaffolds with excellent biocompatibility and biodegradability by electrospinning technology and a chemical cross-linking method in this study. The effect of electrospinning parameters on the structure and morphology, the mechanism of chemical cross-linking, biocompatibility in vitro cell culture experiments, and biodegradability in phosphate-buffered saline buffer solution and trypsin solution of the γ-PGA/keratin electrospun nanofibrous scaffolds (ENS) was studied. The results show that the cross-linked γ-PGA/keratin ENSs had excellent water stability and biodegradability. The γ-PGA/keratin ENSs showed better biocompatibility in promoting cell adhesion and cell growth compared with the γ-PGA ENSs. It is expected that γ-PGA/keratin ENSs will be easily and significantly used in tissue engineering to repair or regenerate materials.

Microbially produced gamma poly glutamic acid (γ–PGA) is a commercially important biopolymer with many applications in foods and various other substances and are abundantly used in different parts of the world. With an aim to study the potent γ–PGA producing Bacillus species, a total of 47 different samples (Kinema, soil, and water) were randomly collected from different locations across the country, and Bacillus sp. were selectively isolated, screened, and characterized by performing physiological, biochemical, morphological, and 16S rRNA gene sequencing. The microbial production of γ–PGA was assayed with the selected isolates on the PGA medium and the metabolite obtained was recovered by ethanol precipitation method and further characterized by thin-layer chromatography (TLC). Thermotolerance (25–60 °C), pH tolerance (4–9), and NaCl tolerance (1–9%) tests were performed to optimize the bacterial growth and γ–PGA production and its viscosity were measured by Ostwald's viscometer. Out of 145 randomly selected colonies, 63 isolates were Gram-positive, rods, and endospore producers and were presumptively confirmed as genus Bacillus. Higher growth of γ–PGA producers were reported in 22 isolates and was found at optimum conditions such as temperature (30–37 °C), pH (6.5–7), incubation time (3 days), and NaCl concentration (3%) and γ–PGA thus produced was further verified by TLC with the retention factor (RF) value 0.27. The potent isolates were closely similar to Bacillus subtilis subsp. stercoris, Bacillus cereus, Bacillus paranthracis, and Bacillus licheniformis etc. Based on the findings of the study, B. licheniformis is the most potent γ–PGA producing Bacillus sp. which can further be used for the commercial production of γ–PGA. To the best of our knowledge, there is yet no published research from Nepal showing the production of the γ–PGA although microbially produced γ–PGA are the major constituents in some popular foods in particular communities of the country.

Casein-based hydrogels were reported as biodegradability, biocompatibility, and non-toxic materials that had potential in drug delivery. At present, we prepared two kinds of casein/γ-PGA hybrid hydrogels, 1/5 and 1/9, based on the ratio of γ-PGA to casein. The hydrogels were crosslinked by microbial transglutaminase (MTG), the physicochemical properties of the casein/γ-PGA hydrogels were investigated by scanning electron microscopy (SEM) observation, differential scanning calorimetry (DSC) analysis, texture analysis, swelling ratio test, and stability test. The hydrogels showed a well-interconnected sparse and porous structure. The 1/5 casein/γ-PGA hydrogel was much stable, hard, and cohesive than the 1/9 casein/γ-PGA hydrogel, and the 1/5 casein/γ-PGA hydrogel showed a higher swelling ratio and lower degradation rate. To investigate in vitro release behavior, we chose the hydrophilic vitamin B12 and hydrophobic aspirin as the model drugs incorporated into the casein/γ-PGA hydrogels. The 1/5 casein/γ-PGA hydrogel exhibited a good drug release behavior. [ABSTRACT FROM AUTHOR]

Presently, there have been some limitations in most of methods to determine poly (γ-glutamic acid) (γ-PGA) content because of many impurities in test specimens. It is necessary to establish a rapid and accurate method to quantify γ-PGA content. In this work, γ-PGA and some impurities commonly seen in fermented broth like glucose, glutamic acid and proteins were used to complex with copper ions. The results show that only γ-PGA can make copper ion precipitated, which content linearly correlates with the precipitate amount. From the study on the validity of the method, it is found that the accuracy and precision are 95.82% and 99.29%, much higher than the ones of method UV and weighing. Therefore, the method via the complexation of copper ion will be popularized to determine γ-PGA content in crude biological samples. • Poly-(γ-glutamic acid) (γ-PGA) can complex with copper ion. • A rapid and accurate method to quantify γ-PGA content in crude sample • The method can determine high concentration of γ-PGA sample. [ABSTRACT FROM AUTHOR]

Three genes involved in poly-γ-glutamic acid(γ-PGA)synthesis cloned from Bacillus licheniformis were transformed into cucumber for the first time. Compared with control, its water content increased by 6–14 % and water loss rate decreased by 11–12 %. In zebrafish and human skin experiments, the moisturizing effect of transgenic cucumber was significantly higher than that of CK, γ-PGA and hyaluronic acid group. Transgenic cucumber reduced facial wrinkles and roughness by 19.58 % and 24.97 %, reduced skin melanin content by 5.27 %, increased skin topological angle and L-value by 5.89 % and 2.49 %, and increased the R2 and Q1 values of facial elasticity by 7.67 % and 5.64 %, respectively. The expressions of aqp3 , Tyr , silv and OCA2 were down-regulated, eln1 , eln2 , col1a1a and col1a1b were up-regulated in zebrafish after treated with transgenic cucumber. This study provides an important reference for the endogenous synthesis of important skin care functional molecules in plants. The transgenic cucumber could downregulate the expression of aqp3 that was lead to increasing by the addition of NaCl, prevent the formation of "pores" on the cell membrane, control the water in cells, and lock moisture in the skin; also, it could maintain skin elasticity through upregulating the expression of collagen genes col1a1a and col1a1b and the elastin genes eln1 and eln2 ; further, it could improve skin tone through downregulating the genes of tyr , silv and oca2 in the process of the formation of melanin. [Display omitted] • Cucumber successfully synthesized γ-PGA heterologously, and the normal growth of cucumber was not affected. • Its water content increased by 6~14% and water loss rate decreased by 11-12% owing to the heterologous γ-PGA. • Its skin-moisturizing, anti-wrinkle, and whitening effects surpassed those of wild-type and γ-PGA controls significantly. • The transgenic cucumber's skin moisturizing, anti-wrinkle, and whitening enhancement mechanism was confirmed in zebrafish. [ABSTRACT FROM AUTHOR]