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There is a renewed interest in acetone-butanol-ethanol (ABE) fermentation from renewable substrates for the sustainable and environment-friendly production of biofuel and platform chemicals. However, the ABE fermentation is associated with several challenges due to the presence of heterogeneous components in the renewable substrates and the intrinsic characteristics of ABE fermentation process. Hence, there is a need to select optimal substrates and modify their characteristics suitable for the ABE fermentation process or microbial strain. This "designed biomass" can be used to establish the consolidated bioprocessing systems. As there are very few reports on designed biomass, the main objectives of this review are to summarize the main challenges associated with ABE fermentation from renewable substrates and to introduce feasible strategies for designing the substrates through pretreatment and hydrolysis technologies as well as through the establishment of consolidated bioprocessing systems. This review offers new insights on improving the efficiency of ABE fermentation from designed renewable substrates.

Utilization of lignocellulosic biomasses for biobutanol fermentation usually requires costly processes of pretreatment and enzymatic hydrolysis. In this study, paper pulp (93.2% glucan) was used as a starting biomass material to produce biobutanol. We conducted enzymatic semi-hydrolysis of paper pulp without pretreatment and with low enzyme loading, which produced high concentrations of cellobiose (13.9 g L -1 ) and glucose (21.3 g L -1 ). In addition, efficient fermentation of the semi-hydrolysate was achieved similar to that with the use of commercial sugars without inhibitors. Finally, we designed a novel non-isothermal simultaneous saccharification and fermentation with in situ butanol recovery, which was composed of a repeated semi-hydrolysis process and successive butanol-extractive fermentation process under the respective optimal conditions. The consolidated system improved butanol production, butanol yields, and butanol productivities and enabled repeated use of medium when compared with other integrated hydrolysis and fermentation processes., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Product inhibition by butanol and acetone is a known drawback in acetone-butanol-ethanol (ABE) fermentation. Extractive fermentation improves butanol production by several ABE-producing Clostridium spp., but only low volume ratios (<4) of extractant to broth (V e /V b ) have been studied. Here, a novel extractive fermentation process was developed using Clostridium saccharoperbutylacetonicum N1-4 and a large V e /V b ratio. A mixture of oleyl alcohol-tributyrin (1:1 (v/v)) yielded high distribution coefficients for both butanol (3.14) and acetone (0.660). Although a fed-batch culture using free cells and the oleyl alcohol-tributyrin mixture at a V e /V b ratio of 5 had a lag phase of >24 h, it produced a higher concentration of total butanol (i.e., butanol produced in all the phases per broth volume used) of 24.2 g/L-broth after 96 h compared with 14.4 g/L-broth at a V e /V b ratio of 1, resulting in a low butanol concentration in the aqueous phase. The use of cells immobilized with calcium alginate beads shortened the lag phase to <12 h. Butanol production was achieved not only in a 3-phase mode (extractant, beads, and tryptone-yeast extract-acetate (TYA) medium) but also in a 2-phase mode (extractant and beads containing TYA medium, without an aqueous phase) at a V e /V b ratio of 5, resulting butanol concentrations of 30.9 g/L-broth and 27.7 g/L-broth, respectively. The 3-phases fed-batch extractive fermentation at a V e /V b ratio of 10 showed a better performance compared with published reports: a total butanol concentration of 64.6 g/L-broth and a butanol yield to consumed sugar of 0.378 C-mol/C-mol., (Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

To improve butanol fermentation efficiencies, semi-hydrolysate with low enzyme loading using H 2 SO 4 pretreated rice straw was designed, which preferably produced cellobiose with xylose (instead of glucose). Fermentation of semi-hydrolysates avoided carbon catabolite repression (CCR) and produced higher butanol yield to enzyme loading (0.0290 g U -1 ), a newly proposed parameter, than the conventional glucose-oriented hydrolysate (0.00197 g U -1 ). Further, overall butanol productivity was improved from 0.0628 g L -1  h -1 to 0.265 g L -1  h -1 during fermentation of undetoxified semi-hydrolysate by using high cell density. A novel simultaneously repeated hydrolysis and fermentation (SRHF) was constructed by recycling of enzymes and cells, which further improved butanol yield to enzyme loading by 183% and overall butanol productivity by 6.04%. Thus, semi-hydrolysate with SRHF is a smartly designed biomass for efficient butanol fermentation of lignocellulosic materials., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Lactic acid (LA) fermentation requires a neutralizer for a physiologically acceptable range. However, a neutralizer generates a large amount of gypsum, an environmental pollutant. Furthermore, the downstream processing is complicated and expensive, comprising 50-70% of the total cost. We previously developed a Lactobacillus delbrueckii FM1, which can produce undissociated LA without neutralizer. Here, we improved FM1 by adaptive evolution at pH 4.5, which generated Adp FM1 showing an ~ 1.80-fold increase in LA production compared to FM1. The LA production via fed-batch fermentation yielded 36.2 g/L of LA, with a productivity of 0.500 g/L/h. However, cell viability was reduced due to the acidic pH and/or end-product inhibition. Therefore, an in situ LA recovery process using an extractive solvent was employed to maintain cell viability. Adp FM1 produced 49.2 g/L of LA via in situ LA-extractive fed-batch fermentation, which was ~ 1.4-fold higher than that without LA extraction. Adp FM1 provided a total LA productivity of 0.512 g/L/h in 96 h. Among the tested strains, Adp FM1 exhibited the highest H + -ATPase activity and a 415-fold increase in H + -ATPase gene expression compared to the parent strain. These results suggest that the in situ LA extractive fermentation process will ease downstream processing and prove to be a more economical and environmentally friendly option compared to the present fermentation. To our knowledge, this is the first report on the production of undissociated L-LA by Lactobacillus using an in situ recovery process, with high LA production levels and productivity.

Lactic acid (LA) is an important platform chemical due to its significant applications in various fields and its use as a monomer for the production of biodegradable poly(lactic acid) (PLA). Free LA production is required to get rid of CaSO 4 , a waste material produced during fermentation at neutral pH which will lead to easy purification of LA required for the production of biodegradable PLA. Additionally, there is no need to use corrosive acids to release free LA from the calcium lactate produced during neutral fermentation. To date, several attempts have been made to improve the acid tolerance of lactic acid bacteria (LAB) by using both genome-shuffling approaches and rational design based on known mechanisms of LA tolerance and gene deletion in yeast strains. However, the lack of knowledge and the complexity of acid-tolerance mechanisms have made it challenging to generate LA-tolerant strains by simply modifying few target genes. Currently, adaptive evolution has proven an efficient strategy to improve the LA tolerance of individual/engineered strains. The main objectives of this article are to summarize the conventional biotechnological LA fermentation processes to date, assess their overall economic and environmental cost, and to introduce modern LA fermentation strategies for free LA production. In this review, we provide a broad overview of free LA fermentation processes using robust LAB that can ferment in acidic environments, the obstacles to these processes and their possible solutions, and the impact on future development of free LA fermentation processes commercially.

Stillage reflux fermentation in food waste ethanol fermentation could reduce sewage discharge but exert a harmful effect because of side-product accumulation. In this study, regulation methods based on metabolic regulation and side-product alleviation were conducted. Result demonstrated that controlling the proper oxidation-reduction potential value (-150mV to -250mV) could reduce the harmful effect, improve ethanol yield by 21%, and reduce fermentation time by 20%. The methods of adding calcium carbonate to adjust the accumulated lactic acid showed that ethanol yield increased by 17.3%, and fermentation time decreased by 20%. The accumulated glyceal also shows that these two methods can reduce the harmful effect. Fermentation time lasted for seven times without effect, and metabolic regulation had a better effect than side-product regulation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Raw materials and pollution control are key issues for the ethanol fermentation industry. To address these concerns, food waste was selected as fermentation substrate, and stillage reflux was carried out in this study. Reflux was used seven times during fermentation. Corresponding ethanol and reducing sugar were detected. Accumulation of by-products, such as organic acid, sodium chloride, and glycerol, was investigated. Lactic acid was observed to accumulate up to 120g/L, and sodium chloride reached 0.14mol/L. Other by-products did not accumulate. The first five cycles of reflux increased ethanol concentration, which prolonged fermentation time. Further increases in reflux time negatively influenced ethanol fermentation. Single-factor analysis with lactic acid and sodium chloride demonstrated that both factors affected ethanol fermentation, but lactic acid induced more effects., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Butanol has recently attracted attention as an alternative biofuel because of its various advantages over other biofuels. Many researchers have focused on butanol fermentation with renewable and sustainable resources, especially lignocellulosic materials, which has provided significant progress in butanol fermentation. However, there are still some drawbacks in butanol fermentation in terms of low butanol concentration and productivity, high cost of feedstock and product inhibition, which makes butanol fermentation less competitive than the production of other biofuels. These hurdles are being resolved in several ways. Genetic engineering is now available for improving butanol yield and butanol ratio through overexpression, knock out/down, and insertion of genes encoding key enzymes in the metabolic pathway of butanol fermentation. In addition, there are also many strategies to improve fermentation technology, such as multi-stage continuous fermentation, continuous fermentation integrated with immobilization and cell recycling, and the inclusion of additional organic acids or electron carriers to change metabolic flux. This review focuses on the most recent advances in butanol fermentation especially from the perspectives of genetic engineering and fermentation technology., (Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The development and implementation of renewable materials for the production of versatile chemical resources have gained considerable attention recently, as this offers an alternative to the environmental problems caused by the petroleum industry and the limited supply of fossil resources. Therefore, the concept of utilizing biomass or wastes from agricultural and industrial residues to produce useful chemical products has been widely accepted. Lactic acid plays an important role due to its versatile application in the food, medical, and cosmetics industries and as a potential raw material for the manufacture of biodegradable plastics. Currently, the fermentative production of optically pure lactic acid has increased because of the prospects of environmental friendliness and cost-effectiveness. In order to produce lactic acid with high yield and optical purity, many studies focus on wild microorganisms and metabolically engineered strains. This article reviews the most recent advances in the biotechnological production of lactic acid mainly by lactic acid bacteria, and discusses the feasibility and potential of various processes., (Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Nukadoko is a fermented rice bran mash traditionally used for pickling vegetables in Japan. To date, the production of both homemade and commercial nukadoko depends on natural fermentation without using starter cultures. Here, we monitored chemical and microbiological changes in the initial batch fermentation of nukadoko. Nukadoko samples were prepared by spontaneous fermentation of four different brands of rice bran, and microbiome dynamics were analyzed for 2 months. In the first week, non-Lactobacillales lactic acid bacteria (LAB) species, which differed among the samples, grew proportionally to pH decrease and lactate increase. Thereafter, Lactobacillus plantarum started growing and consumed residual sugars, causing further lactate increase in nukadoko. Finally, microbial communities in all tested nukadoko samples were dominated by L. plantarum. Taken together, our results suggest that the mixture of the fast-growing LAB species and slow-growing L. plantarum may be used as a suitable starter culture to promote the initial fermentation of nukadoko., (Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Fermentative production of optically pure lactic acid has roused interest among researchers in recent years due to its high potential for applications in a wide range of fields. More specifically, the sharp increase in manufacturing of biodegradable polylactic acid (PLA) materials, green alternatives to petroleum-derived plastics, has significantly increased the global interest in lactic acid production. However, higher production costs have hindered the large-scale application of PLA because of the high price of lactic acid. Therefore, reduction of lactic acid production cost through utilization of inexpensive substrates and improvement of lactic acid production and productivity has become an important goal. Various methods have been employed for enhanced lactic acid production, including several bioprocess techniques facilitated by wild-type and/or engineered microbes. In this review, we will discuss lactic acid producers with relation to their fermentation characteristics and metabolism. Inexpensive fermentative substrates, such as dairy products, food and agro-industrial wastes, glycerol, and algal biomass alternatives to costly pure sugars and food crops are introduced. The operational modes and fermentation methods that have been recently reported to improve lactic acid production in terms of concentrations, yields, and productivities are summarized and compared. High cell density fermentation through immobilization and cell-recycling techniques are also addressed. Finally, advances in recovery processes and concluding remarks on the future outlook of lactic acid production are presented., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Nukadoko is a naturally fermented rice bran mash traditionally used for pickling vegetables in Japan; its refreshment and fermentation cycles sometimes continue for many years. Here, we investigated the structure and dynamics of the bacterial community in nukadoko by conducting pyrosequencing and quantitative polymerase chain reaction (PCR) analyses of 16S ribosomal RNA genes (rDNA). Of the 16 different samples studied, 13 showed Lactobacillus-dominated microbiota, suggesting that aged nukadoko samples tend to realize a niche, favorable Lactobacillus species. The lactic acid bacterial community of each of the 16 samples was classified into 3 types according to the presence or absence of 2 predominant species, Lactobacillus namurensis and Lactobacillus acetotolerans. The dynamics of the bacterial community during fermentation and the subsequent ripening process were examined using a laboratory model of nukadoko inoculated with an aged nukadoko sample (inoculated model). Lb. namurensis grew rapidly in the first 2 days, accompanied with a rapid decrease in pH and an increase in lactate levels, while Lb. acetotolerans grew with a longer doubling time and slow acidification during the 20 days after inoculation. On the other hand, spontaneous fermentation of the nukadoko model prepared from fresh rice bran without the nukadoko inoculation (inoculant-free model), showed the growth of some non-Lactobacillus species such as staphylococci and bacilli within the first 10 days; thereafter, Lb. namurensis was dominant, while Lb. acetotolerans was not detected during the 20-day experimental period. These results suggest that the naturally established Lactobacillus community in aged nukadoko is effectively involved in the biocontrol of the microbial community of nukadoko during the refreshment and fermentation cycles., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Fermented barley extract (FBE) obtained from a barley shochu by-product (shochu kasu) and its ethanol fractions were evaluated as a medium and supplement, respectively, for nisin A production by Lactococcus lactis subsp. lactis ATCC 11454. A Brix 2.5 FBE medium supplemented with glucose provided a high level of nisin A production with a nisin yield comparable to that from a nutritionally rich laboratory medium (basal medium). By adding the ethanol-insoluble (EI) fraction of FBE to the basal medium, nisin A production was enhanced concomitant with an increase in bacterial cell growth, while the ethanol-soluble (ES) fraction had a negative effect on nisin A production. These findings indicate that FBE obtained from shochu kasu can be utilized as a preferable medium for nisin A production and could be converted into a value-added food product having preservative functions. The procedure developed in this study would promote recycling of shochu kasu.

Background: The simultaneous and effective conversion of both pentose and hexose in fermentation is a critical and challenging task toward the lignocellulosic economy. This study aims to investigate the feasibility of an innovative co-fermentation process featuring with a cell recycling unit (CF/CR) for mixed sugar utilization. A l-lactic acid-producing strain Enterococcus mundtii QU 25 was applied in the continuous fermentation process, and the mixed sugars were utilized at different productivities after the flowing conditions were changed. A mathematical model was constructed with the experiments to optimize the biological process and clarify the cell metabolism through kinetics analysis. The structured model, kinetic parameters, and achievement of the fermentation strategy shall provide new insights toward whole sugar fermentation via real-time monitoring for process control and optimization. Results: Significant carbon catabolite repression in co-fermentation using a glucose/xylose mixture was overcome by replacing glucose with cellobiose, and the ratio of consumed pentose to consumed hexose increased significantly from 0.096 to 0.461 by mass. An outstanding product concentration of 65.2 g L−1 and productivity of 13.03 g L−1 h−1 were achieved with 50 g L−1 cellobiose and 30 g L−1 xylose at an optimized dilution rate of 0.2 h−1, and the cell retention time gradually increased. Among the total lactic acid production, xylose contributed to more than 34% of the mixed sugars, which was close to the related contents in agricultural residuals. The model successfully simulated the transition of sugar consumption, cell growth, and lactic acid production among the batch, continuous process, and CF/CR systems. Conclusion: Cell retention time played a critical role in balancing pentose and hexose consumption, cell decay, and lactic acid production in the CF/CR process. With increasing cell concentration, consumption of mixed sugars increased with the productivity of the final product; hence, the impact of substrate inhibition was reduced. With the validated parameters, the model showed the highest accuracy simulating the CF/CR process, and significantly longer cell retention times compared to hydraulic retention time were tested. [ABSTRACT FROM AUTHOR]

Presently, several researchers are increasingly focusing on producing butanol as the next-generation fuel by acetone-butanol-ethanol ( ABE) fermentation. Butanol has many superior characteristics compared to other biofuels, such as ethanol. However, its production by ABE fermentation faces the challenges of low productivity and yield because of product inhibition and heterofermentation, respectively, and thereby, high costs. Until date, molecular biological techniques and fermentation engineering methods have been applied for high butanol production. Although glucose remains the substrate of choice since traditional research, it is now necessary to substitute glucose derived from edible starch to other substrates from low-cost feedstock, such as agricultural residue. In addition, ABE-producing clostridia cannot directly produce butanol from lignocelluloses. Therefore, recent research is focusing on pretreatment and enzymatic saccharification of the complex molecules derived from agricultural residue for use as feedstock in butanol production. This article reviews traditional research, including the metabolism patterns and characteristics of ABE-producing clostridia. Furthermore, this article describes developments in ABE fermentation with respect to the establishment of highly efficient butanol production processes, such as batch, fed-batch, and continuous cultures, with the introduction of butanol removal, as well as butanol production from lignocellulosic biomasses or alternative substrates to sugars. [ABSTRACT FROM AUTHOR]

Abstract: The conventional continuous fermentation and continuous with high cell density for lactic acid production from cassava starch by Lactobacillus plantarum SW14 was investigated. In conventional continuous fermentation, lactic acid productivity increased while the acid concentration decreased with increasing dilution rate. The highest lactic acid productivity of 4.53 gl-1h-1 was observed at a dilution rate of 0.4 h-1, which was 4.7 time higher than batch fermentation. A further increase in lactic acid productivity was observed in a continuous with high cell density and reached 8.0 gl-1h-1, which were equal to 8 and 1.8 times compared to batch and conventional continuous mode, respectively. These results indicated thatL. plantarum SW14 has a potential for lactic acid production directly from cassava starch when continuous mode was applied with high cell density and operated at high dilution rate. [Copyright &y& Elsevier]

Effective utilisation of cellulosic biomasses for economical lactic acid production requires a microorganism with potential ability to utilise efficiently its major components, glucose and cellobiose. Amongst 631 strains isolated from different environmental samples, strain QU 25 produced high yields of l-(+)-lactic acid of high optical purity from cellobiose. The QU 25 strain was identified as Enterococcus mundtii based on its sugar fermentation pattern and 16S rDNA sequence. The production of lactate by fermentation was optimised for the E. mundtii QU25 strain. The optimal pH and temperature for batch culturing were found to be 7.0°C and 43°C, respectively. E. mundtii QU 25 was able to metabolise a mixture of glucose and cellobiose simultaneously without apparent carbon catabolite repression. Moreover, under the optimised culture conditions, production of optically pure l-lactic acid (99.9%) increased with increasing cellobiose concentrations. This indicates that E. mundtii QU 25 is a potential candidate for effective lactic acid production from cellulosic hydrolysate materials. [ABSTRACT FROM AUTHOR]

In order to achieve high butanol production by Clostridium saccharoperbutylacetonicum N1-4, the effect of lactic acid on acetone–butanol–ethanol fermentation and several fed-batch cultures in which lactic acid is fed have been investigated. When a medium containing 20 g/l glucose was supplemented with 5 g/l of closely racemic lactic acid, both the concentration and yield of butanol increased; however, supplementation with more than 10 g/l lactic acid did not increase the butanol concentration. It was found that when fed a mixture of lactic acid and glucose, the final concentration of butanol produced by a fed-batch culture was greater than that produced by a batch culture. In addition, a pH-controlled fed-batch culture resulted in not only acceleration of lactic acid consumption but also a further increase in butanol production. Finally, we obtained 15.5 g/l butanol at a production rate of 1.76 g/l/h using a fed-batch culture with a pH-stat continuous lactic acid and glucose feeding method. To confirm whether lactic acid was converted to butanol by the N1-4 strain, we performed gas chromatography–mass spectroscopy (GC-MS) analysis of butanol produced by a batch culture during fermentation in a medium containing [1,2,3-13C3] lactic acid as the initial substrate. The results of the GC-MS analysis confirmed the bioconversion of lactic acid to butanol. [ABSTRACT FROM AUTHOR]

An amendment to this paper has been published and can be accessed via the original article. [ABSTRACT FROM AUTHOR]

Eucalyptus has been previously suggested as a potential substrate for acetone-butanol-ethanol (ABE) production without nutrient supplementation; however, incomplete sugar consumption has prevented improvement of ABE production. Cellulase loading with 35 FPU g −1 was first optimized in terms of high hydrolysis efficiency (95%). However, only 0.43 g L −1 ABE production and 3.44% glucose consumption rate were achieved. To improve ABE production from eucalyptus hydrolysate, supplementation of 6 nutrients in common tryptone-yeast extract (TY) medium were investigated by statistical approaches. Three nutrients including yeast extract, tryptone, and FeSO 4 ·7H 2 O were screened as significant nutrients for ABE production. Subsequently, use of a modified TY medium (MTY medium: yeast extract 3.04 g L −1 , tryptone 7.64 g L −1 , FeSO 4 ·7H 2 O 15.3 mg L −1 ), which was subsequently predicted by Plackett-Burman and Box-Behnken designs to stimulate ABE production, resulted in ca. 40-fold increase in ABE concentration (16.9 g L −1 ) and a glucose consumption rate of 100%. We first examined previously uninvestigated nutrition combinations using Plackett-Burman and Box-Behnken designs for high ABE production from eucalyptus hydrolysate. This study shows that statistical method would be a powerful tool for the optimization and enhancement of ABE production from eucalyptus hydrolysate. [ABSTRACT FROM AUTHOR]

To reuse organic acids in wastewater, efficient acetone-butanol-ethanol (ABE) fermentation was constructed using mixed acetic acid and lactic acid. The mixed acids were co-consumed with glucose, producing higher butanol concentration (6.96 g/L) and butanol yield (0.494 C-mol/C-mol) than using each single acid (6.22 g/L and 0.432 C-mol/C-mol). This result was verified by 13C tracer experiment that addition of lactic acid improved carbon distribution to butanol instead of acetone, while acetic acid was readily consumed than lactic acid at initial fermentation stage. Subsequently, pH-stat fed batch fermentation at low pH of 5.5 further improved acid consumptions (6.34 g/L acetic acid and 9.06 g/L lactic acid) and solvent productions (12.4 g/L butanol and 6.50 g/L acetone). Finally, ABE fermentation using wastewater of lactic acid fermentation was performed without inhibition. Thus, efficient ABE fermentation can be implemented using mixed acetic acid and lactic acid, which is superior to using single acid. • This is the first study using mixed acetate and lactate for ABE fermentation. • 13C experiment verified the metabolic flux to butanol was improved by the acids. • pH and glucose played an important role in acid consumption and solvent production. • Fermentation from lactic acid fermentation broth was performed without inhibition. [ABSTRACT FROM AUTHOR]

We previously reported a butanol production process with pH-stat continuous feeding of dl-lactic acid and glucose as the co-substrate (Oshiro et al., Appl. Microbiol. Biotechnol., 87, 1177–1185, 2010). To accomplish butanol production from completely inedible substrates, in this study, we investigated acetone-butanol-ethanol (ABE) fermentation of Clostridium saccharoperbutylacetonicum N1-4 with lactic acid by using pentose as the co-substrate. Examination for optimum co-substrate indicated that arabinose was superior to glucose and xylose for ABE fermentation. Actually batch culture with lactic acid and arabinose without pH control exhibited higher butanol production (7.11 g/l) and lactic acid consumption (2.02 g) than those (6.62 g/l and 1.45 g, respectively) with glucose. Fed-batch culture without pH control increased these values to 12.08 g/l and 15.60 g/l butanol production, and to 3.83 g and 5.91 g lactic acid consumption by feeding the substrate once and twice, respectively. Finally, the result of gas chromatography–mass spectroscopy analysis using [1,2,3-13C3]-lactic acid indicated that lactic acid was converted to butanol with the efficiency of 51.9%. Thus, we established a novel high butanol production from lactic acid using arabinose as the co-substrate in simple fed-batch culture. [ABSTRACT FROM AUTHOR]

Abstract: Lactic acid is an industrially important product with a large and rapidly expanding market due to its attractive and valuable multi-function properties. The economics of lactic acid production by fermentation is dependent on many factors, of which the cost of the raw materials is very significant. It is very expensive when sugars, e.g., glucose, sucrose, starch, etc., are used as the feedstock for lactic acid production. Therefore, lignocellulosic biomass is a promising feedstock for lactic acid production considering its great availability, sustainability, and low cost compared to refined sugars. Despite these advantages, the commercial use of lignocellulose for lactic acid production is still problematic. This review describes the “conventional” processes for producing lactic acid from lignocellulosic materials with lactic acid bacteria. These processes include: pretreatment of the biomass, enzyme hydrolysis to obtain fermentable sugars, fermentation technologies, and separation and purification of lactic acid. In addition, the difficulties associated with using this biomass for lactic acid production are especially introduced and several key properties that should be targeted for low-cost and advanced fermentation processes are pointed out. We also discuss the metabolism of lignocellulose-derived sugars by lactic acid bacteria. [Copyright &y& Elsevier]

Recently, lactic acid bacteria (LAB) have attracted much attention because of their potential application to probiotics and industrial applications as starters for dairy products or lactic acid fermentation. Additional emphasis is also being paid to them as commensal bacteria in gastrointestinal tract. Since LAB exhibit a stress response, insight into the relationship between stress proteins such as molecular chaperones and stress tolerance or adaptation is increasing gradually along with current research examining these important bacteria. Similar to other bacteria, one of the major stress-response systems in LAB is the expression of molecular chaperones. The recently completed genome sequencing of various LAB strains, combined with the development of advanced molecular techniques, have enabled us to identify molecular chaperones and to understand their regulation systems in response to various stresses. Furthermore, recent biochemical studies provided novel insight into the molecular mechanisms of LAB chaperone systems. This review highlights the physiological consequences and biochemical properties of molecular chaperones (especially sHsps, Hsp70, and Hsp100) in LAB and their use in biotechnological applications. [Copyright &y& Elsevier]

Strains of Leuconostoc lactis SHO-47 and Le. lactis SHO-54, producing the clinically useful enzyme NAD-specific 6-phosphoglucanate dehydrogenase, were cultivated with a hydrolyzed birch wood xylan as the unique carbon source to produce D-lactic acid for poly(D-lactic acid). In addition to the strains SHO-47 and SHO-54, Lactococcus lactis IO-1, well known as a good xylose-utilizing lactic acid bacterium, was used as a control to confirm the extent of hemicellulose hydrolysis. The fermentation time for lactic acid of strains SHO-47 and SHO-54 was 12 h, and produced respectively 2.3 and 2.2 g/l lactic acid from 8.5 g/l hydrolyzed xylan, whereas the fermentation time of strain IO-1 was 21 h, and produced 1.3 g/l lactic acid. Xylooligosaccharides from xylobiose to xylohexose were utilized more rapidly than xylose in the cultures of strains SHO-47 and SHO-54. However, xylose concentration increased temporarily and then decreased in the culture of strain IO-1. On the other hand, xylooligosaccharides larger than xyloheptaose were not utilized by these three strains. The xylosidase activities of SHO-47, SHO-54, and IO-1 were induced by xylose or a mixture of xylobiose and xylotriose. The xylosidases of these three strains were localized in their cytoplasm. [Copyright &y& Elsevier]

There has been tremendous growth in the production of optically pure l -lactic acid from lignocellulose-derived sugars. In this study, Enterococcus mundtii QU 25 was used to ferment a glucose/xylose mixture to l -lactic acid. Maintenance of the xylose concentration at greater than 10 g/L achieved homo-lactic acid fermentation and reduced the formation of byproducts. Furthermore, carbon catabolite repression (CCR) was avoided by maintaining the glucose concentration below 25 g/L; therefore, initial concentrations of 25 g/L glucose and 50 g/L xylose were selected. Supplementation with 5 g/L yeast extract enhanced the maximum xylose consumption rate and consequently increased lactic acid production and productivity. Finally, a 129 g/L lactic acid without byproducts was obtained with a maximum lactic acid productivity of 5.60 g/(L·h) in fed-batch fermentation with feeding a glucose/xylose mixture using ammonium hydroxide as the neutralizing agent. These results indicate a potential for lactic acid production from glucose and xylose as the main components of lignocellulosic biomasses. [ABSTRACT FROM AUTHOR]

Acetone–butanol–ethanol (ABE) fermentation using Clostridium saccharoperbutylacetonicum N1-4 and mixed sugars containing cellobiose and xylose was studied to establish efficient butanol production process without carbon catabolite repression (CCR). Although batch culture with glucose and xylose exhibited apparent CCR, we achieved simultaneous consumption of cellobiose and xylose. Moreover, preculture of the N1-4 strain with xylose yielded maximum butanol and solvent concentrations (16 and 23 g/L, respectively). Thus, we succeeded in ABE fermentation with mixed sugars of hexose and pentose, without CCR, by using wild-type ABE-producing clostridia. We also investigated the effect of various ratios of cellobiose and xylose on the fermentation process and yield. Increasing initial xylose concentration improved butanol and solvent concentrations and maximum xylose consumption rate. Fed-batch culture with cellobiose and xylose showed rapid and simultaneous sugar consumption and improved maximum consumption rate of both sugars. [ABSTRACT FROM AUTHOR]

Abstract: A continuous butanol production system with high-density Clostridium saccharoperbutylacetonicum N1-4 generated by cell recycling was established to examine the characteristics of butanol fermentation from xylose. In continuous culture without cell recycling, cell washout was avoided by maintaining pH>5.6 at a dilution rate of 0.26h−1, indicating pH control was critical to this experiment. Subsequently, continuous culture with cell recycling increased cell concentration to 17.4gL−1, which increased butanol productivity to 1.20gL−1 h−1 at a dilution rate of 0.26h−1 from 0.529gL−1 h−1 without cell recycling. The effect of dilution rates on butanol production was also investigated in continuous culture with cell recycling. Maximum butanol productivity (3.32gL−1 h−1) was observed at a dilution rate of 0.78h−1, approximately 6-fold higher than observed in continuous culture without cell recycling (0.529gL−1 h−1). [Copyright &y& Elsevier]

Enterococcus faecium NKR-5-3 produces four antimicrobial peptides referred here as enterocins NKR-5-3A, B, C and D. A two-step electrospray ionization-liquid chromatography and mass spectrometry (ESI-LC/MS)-based quantification system was developed to monitor its multiple bacteriocin production profiles, which is essential in understanding the complex production regulation mechanism of strain NKR-5-3. The developed ESI-LC/MS-based quantification system can easily monitor the multiple bacteriocin production of this strain. Using the developed system, the production of enterocin NKR-5-3B was found to be not as variable as those of the other enterocins in different cultivation media. Production of enterocin NKR-5-3B was also found to have a wider optimum incubation temperature (20–30°C) than enterocins NKR-5-3A, C and D (25°C). Furthermore, at least 2 nM of the bacteriocin-like inducing peptide, enterocin NKR-5-3D, regulated the production of NKR-5-3 enterocins except enterocin NKR-5-3B. These findings taken together suggest that enterocin NKR-5-3B has an independent production regulation mechanism from the other NKR-5-3 enterocins. The developed system could effectively pin-point the production profiles of the multiple bacteriocins of E. faecium NKR-5-3 under different fermentation conditions. [ABSTRACT FROM AUTHOR]

Pretreatment of vinasse is necessary to break down its complex wooden structure, improve the efficiency of enzymolysis, and provide an adequate substrate for fermentation during l-lactic acid production. Independent microwave and NaOH pretreatment processes were carried out in an earlier study. Here a two-way microwave-NaOH coupled method, was investigated to determine the effects of simultaneous saccharification and fermentation (SSF) of vinasse on l-lactic acid production. Compared with the former individual microwave and NaOH pretreatment methods, hemicellulose and lignin in vinasse were more significantly reduced using microwave-NaOH coupled pretreatments, reducing crystallinity to 14.77%. l-lactic acid yield was also higher in the coupled method than the sum of the yields from the independent pretreatment processes. The optimal conditions for the coupled pretreatment method were: 0.06 g g−1 (vinasse dry weight) NaOH; 523 W microwave power; 1:2 solid–liquid ratio; and 8 min microwave pretreatment time. These conditions resulted in an 86.0 mg g−1 l-lactic acid yield. The microwave power and the amount of alkali used were approximately 25% lower than those used prior to the optimisation process. The reduced values indicate that optimisation can significantly decrease energy and reagent consumptions. Thus, microwave-NaOH coupled pretreatment exhibits a synergistic effect for improving l-lactic acid production. [Copyright &y& Elsevier]

Abstract: Lactic acid bacteria forming bacteriocin active against Listeria monocytogenes were screened for potential use in controlling growth of L. monocytogenes in Nham, a Thai traditional fermented pork sausage. Based on the spot-on-lawn assay, Pediococcus pentosaceus BCC 3772 was found to produce the highest anti-listeria activity. Through a series of chromatographic techniques, Edman N-terminal sequencing and mass spectrometry, this anti-listeria compound showed a complete homology to pediocin PA-1/AcH. Inoculation of P. pentosaceus BCC 3772 in Nham caused a significant decrease of 3.2 log in population of spiked L. monocytogenes within 18–24 h of fermentation, in comparison with the initial count with no significant changes in sensory properties and consumer acceptability on the overall characteristics of the final fermented Nham products. These results indicate that the P. pentosaceus BCC 3772 should be useful as a functional starter culture for control of L. monocytogenes without compromising the unique quality of Nham. [Copyright &y& Elsevier]

Enterococcus mundtii QU 25, a newly isolated lactic acid bacterium, efficiently metabolized xylose into L-lactate. In batch fermentations, the strain produced 964 mM L-(+)-lactate from 691 mM xylose, with a yield of 1.41 mol/mol xylose consumed and an extremely high optical purity of ⩾99.9% without acetate production. [ABSTRACT FROM AUTHOR]

The effects of nisin-induced dnaK expression in Lactococcus lactis were examined, and this expression was shown to improve stress tolerance and lactic acid fermentation efficiency. Using a nisin-inducible expression system, DnaK proteins from L. lactis (DnaKLla) and Escherichia coli (DnaKEco) were produced in L. lactis NZ9000. In comparison to a strain harboring the empty vector pNZ8048 (designated NZ-Vector) and one expressing dnaKLla (designated NZ-LDnaK), the dnaKEco-expressing strain, named NZ-EDnaK, exhibited more tolerance to heat stress at 40°C in GM17 liquid medium. The cell viability of NZ-Vector was reduced 4.6-fold after 6 h of heat treatment. However, NZ-EDnaK showed 13.5-fold increased viability under these conditions, with a very low concentration of DnaKEco production. Although the heterologous expression of did not effect DnaKEco, production, heat treatment increased the DnaKLla level 3.5- and 3.6-fold in NZ-Vector and NZ-EDnaK, respectively. Moreover, NZ-EDnaK showed tolerance to multiple stresses, including 3% NaCl, 5% ethanol, and 0.5% lactic acid (pH 5.47). In CMG medium, the lactate yield and the maximum lactate productivity of NZ-EDnaK were higher than the corresponding values for NZ-Vector at 30°C. Interestingly, at 40°C, these values of NZ-EDnaK were not significantly different from the corresponding values for the control strain at 30°C. Lactate dehydrogenase (LDH) activity was also found to be stable at 40°C in the presence of DnaKEco. These findings suggest that the heterologous expression of dnaKEco enhances the quality control of proteins and enzymes, resulting in improved growth and lactic acid fermentation at high temperature. [ABSTRACT FROM AUTHOR]

Abstract: We proposed a kinetic simulation model of xylose metabolism in Lactococcus lactis IO-1 that describes the dynamic behavior of metabolites using the simulator WinBEST-KIT. This model was developed by comparing the experimental time-course data of metabolites in batch cultures grown in media with initial xylose concentrations of 20.3–57.8 g/l with corresponding calculated data. By introducing the terms of substrate activation, substrate inhibition, and product inhibition, the revised model showed a squared correlation coefficient (r 2) of 0.929 between the experimental time-course of metabolites and the calculated data. Thus, the revised model is assumed to be one of the best candidates for kinetic simulation describing the dynamic behavior of metabolites. Sensitivity analysis revealed that pyruvate flux distribution is important for higher lactate production. To confirm the validity of our kinetic model, the results of the sensitivity analysis were compared with enzyme activities observed during increasing lactate production by adding natural rubber serum powder to the xylose medium. The experimental results on pyruvate flux distribution were consistent with the prediction by sensitivity analysis. [Copyright &y& Elsevier]

Abstract: A kinetic simulation model of acetone–butanol–ethanol (ABE) fermentation of xylose (ModelXYL) was proposed by substituting Embden–Meyerhof–Parnas (EMP) pathway equations in the glucose model (ModelGLC) by pentose phosphate (PP) pathway equations of xylose utilization. We estimated the equation parameters of the PP pathway and set other equation parameters to the same as those in ModelGLC, by which ModelXYL exhibited an r 2 value of 0.901 between the experimental time course of metabolites with initial xylose concentrations ranging from 40.7 to 292mM and the calculated values. The results with the developed model suggested that Clostridium saccharoperbutylacetonicum N1-4 has a robust metabolic network in acid- and solvent-producing pathways. Furthermore, sensitivity analysis revealed that slow substrate utilization would be effective for higher butanol production; this coincided with the experimental results. Therefore, we consider the proposed model to be one of the best kinetic simulation candidates describing the dynamic metabolite behavior in ABE production. [Copyright &y& Elsevier]

Nukadoko is the fermented rice bran bed traditionally used for pickling vegetables in Japan. Here, we investigate the bacterial community structure of nukadoko using several culture-independent methods. Denaturing gradient gel electrophoresis (DGGE) and sequence analysis of V2-V3 16S rRNA gene (16S rDNA) fragments amplified from a long-aged nukadoko bacterial community indicated seven predominant operational taxonomic units (OTUs) closely related to known Lactobacillus species. Phylogenetic analysis of these OTUs indicated a major cluster consisting of six OTUs including a dominant OTU closely related to Lactobacillus acidifarinae and one distinct OTU corresponding to Lactobacillus acetotolerans. L. acetotolerans was commonly detected as a dominant species in samples from different seasons. The succession of microbial community structure in the fermentation and ripening processes was investigated using a laboratory model nukadoko. The L. acidifarinae-like bacteria grew rapidly with a pH decrease in the first few days after inoculation, whereas L. acetotolerans grew slowly and became dominant after one week. Real-time quantitative polymerase chain reaction (Q-PCR) showed that the doubling time of L. acetotolerans was 12 h, while that of total bacteria was 4 h. Real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) targeting 16S rRNA showed a low metabolic activity of L. acetotolerans throughout the fermentation and ripening processes. Fluorescence in situ hybridization (FISH) showed that L. acetotolerans was a dominant bacterium in the ripening period and had a low metabolic activity. These results indicate that the slow-growing L. acetotolerans stably dominated nukadoko microbiota after the L. acidifarinae-like bacteria mainly contributed to the lactic acid fermentation of the rice bran. [Copyright &y& Elsevier]

Abstract: Novel amylolytic lactic acid bacterium from puto, fermented raw rice in Philippine, was isolated and characterized. The strain was identified as Enterococcus faecium No. 78 (BIOTECH 10375) by sugar fermentative test and 16S rDNA sequence analysis. Optimum pH and temperature were 6.5 and 30°C, respectively. Direct l-lactic acid fermentation was carried out with various starches, lactic acid productivity with sago starch being similar to that with glucose. Yield of lactic acid from sago starch was higher than those from glucose and other starches. Strain No. 78 was superior to the other amylolytic lactic acid bacteria so far reported on the direct lactic acid fermentation with starches and produced lactic acid of high optical purity (98.6%). In direct lactic acid fermentation with starch, continuous culture has hardly been reported. Continuous culture system with high cell density of strain No. 78 showed higher lactic acid productivity (3.04gl−1 h−1) than those of batch culture (1.105gl−1 h−1) and conventional continuous culture (1.56gl−1 h−1). Even if the dilution rate was increased up to 0.26h−1, the residual starch concentration was controlled at moderately low level below 3.24gl−1. [Copyright &y& Elsevier]