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Blood pressure (BP) is a known cardiovascular risk factor that is hypothesised to be inversely related to choline intake. A previous study suggested that this association may be more apparent in older adults and may differ according to demographic and health characteristics. The primary study objectives are to investigate the cross-sectional associations of total choline intake with BP (n 843) and prevalent hypertension (n 2113) among USA adults aged ≥ 65 years using the sample from the 2011 to 2014 National Health and Nutrition Examination Survey. Logistic and multiple linear regression models for complex surveys were employed for hypertension status and BP, respectively. Effect modification by sex, race, BMI and comorbidity status were separately investigated using an interaction term. Choline intake interacted with BMI (P-interaction = 0·04) such that choline intake tended to be associated with lower odds of hypertension among people with BMI < 18·5 kg/m2 (OR (95 % CI): 0·64 (0·4, 1·00); P = 0·052). Choline intake was not associated with systolic BP (mean ± sem change per 100 mg of choline: -1·03 ± 0·74 mmHg; P = 0·16). In contrast, its relation to diastolic BP differed by cardiovascular comorbidity (P-interaction = 0·03) with a non-significant (P = 0·13) negative direction of association observed among those who were free of comorbidities and a non-significant (P = 0·26) positive direction observed among those with comorbidities. Collectively, these results suggested that the associations of choline intake with BP levels and hypertension risk among older adults are dependent on other risk factors.

Polylactic acid (PLA) was melt-blended with epoxy resin to study the effects of the reaction on the mechanical and thermal properties of the PLA. The addition of 0.5% (wt/wt) epoxy to PLA increased the maximum tensile strength of PLA (57.5 MPa) to 67 MPa, whereas the 20% epoxy improved the elongation at break to 12%, due to crosslinking caused by the epoxy reaction. The morphology of the PLA/epoxy blends showed epoxy nanoparticle dispersion in the PLA matrix that presented a smooth fracture surface with a high epoxy content. The glass transition temperature of PLA decreased with an increasing epoxy content owing to the partial miscibility between PLA and the epoxy resin. The Vicat softening temperature of the PLA was 59 °C and increased to 64.6 °C for 0.5% epoxy. NMR confirmed the reaction between the -COOH groups of PLA and the epoxy groups of the epoxy resin. This reaction, and partial miscibility of the PLA/epoxy blend, improved the interfacial crosslinking, morphology, thermal properties, and mechanical properties of the blends.

This study aimed to select thermotolerant yeast for bioethanol production from cellulose-rich corncob (CRC) residue. An effective yeast strain was identified as Saccharomyces cerevisiae TC-5. Bioethanol production from CRC residue via separate hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF), and prehydrolysis-SSF (pre-SSF) using this strain were examined at 35-42 °C compared with the use of commercial S. cerevisiae . Temperatures up to 40 °C did not affect ethanol production by TC-5. The ethanol concentration obtained via the commercial S. cerevisiae decreased with increasing temperatures. The highest bioethanol concentrations obtained via SHF, SSF, and pre-SSF at 35-40 °C of strain TC-5 were not significantly different (20.13-21.64 g/L). The SSF process, with the highest ethanol productivity (0.291 g/L/h), was chosen to study the effect of solid loading at 40 °C. A CRC level of 12.5% ( w / v ) via fed-batch SSF resulted in the highest ethanol concentrations of 38.23 g/L. Thereafter, bioethanol production via fed-batch SSF with 12.5% ( w / v ) CRC was performed in 5-L bioreactor. The maximum ethanol concentration and ethanol productivity values were 31.96 g/L and 0.222 g/L/h, respectively. The thermotolerant S. cerevisiae TC-5 is promising yeast for bioethanol production under elevated temperatures via SSF and the use of second-generation substrates.

The (R)-phenylacetylcarbinol (PAC) batch biotransformation kinetics for partially purified Candida tropicalis TISTR 5350 pyruvate decarboxylase (PDC) were determined to validate a comprehensive mathematical model in 250 mL scale with 250 mM phosphate buffer/pH 7.0. PDC could convert initial 100/120 mM benzaldehyde/pyruvate substrates to the statistical significantly highest (p ≤ 0.05) maximum PAC concentration (95.8 ± 0.1 mM) and production rate (0.639 ± 0.001 mM min -1 ). A parameter search strategy aimed at minimizing overall residual sum of square (RSS T ) based on a system of six ordinary differential equations was applied to PAC biotransformation profiles with initial benzaldehyde/pyruvate concentration of 100/120 and 30/36 mM. Ten important biotransformation kinetic parameters were then elucidated including the zeroth order activation rate constant due to phosphate buffer species (k a ) of (9.38 ±  < 0.01) ×  10 -6 % relative PDC activity min -1  mM -1 . The validation of this model to independent biotransformation kinetics with initial benzaldehyde/pyruvate concentration of 50/60 mM resulted in relatively good fitting with RSS T , mean sum of square error (MSE), and coefficient of determination (R 2 ) values of 662, 17.4, and 0.9863, respectively.

The effect of ultrasonication on the antioxidant and antibacterial properties of Ceylon spinach ( Basella alba ) extracts (CE) and the shelf life of chilled pork with CE were studied. The CE were ultrasonicated at different power levels (60-100%) for 10-40 min in an ultrasonic bath with the rise of antioxidant activities ( p ≤ 0.05) proportional to the ultrasonication time. The additional investigation of antibacterial activities showed that the ultrasonicated extracts (100 mg/mL) could inhibit and inactivate Staphylococcus aureus and Escherichia coli with the optimal condition of 80% power for 40 min. For shelf-life testing, fresh pork treated with the ultrasonicated extracts at 100 and 120 mg/mL had lower values of thiobarbituric acid reactive substances (TBARS) than the control (without dipping). For food safety as measured by the total microbial count, the fresh pork dipped with 100-120 mg/mL CE extract could be kept at 0 °C for 7 days, 2 to 3 days longer than control meat at 0 and 4 °C, respectively. A sensory evaluation using a nine-point hedonic scale showed that fresh pork dipped with 100-mg/mL CE extracts was accepted by consumers. It is suggested that CE extracts can be applied in the food industry to enhance the quality and extend the shelf life of meat products.

Curcumin is a phenolic compound derived from turmeric roots ( Curcuma longa L.). This research studied the effects of curcumin extract on the properties of chitosan films. The film characteristics measured included mechanical properties, visual aspects, color parameters, light transmission, moisture content, water solubility, water vapor permeability, infrared spectroscopy, and antioxidant activity. The results suggest that adding curcumin to chitosan-based films increases yellowness and light barriers. Infrared spectroscopy analysis showed interactions between the phenolic compounds of the extract and the chitosan, which may have improved the mechanical properties and reduced the moisture content, water solubility, and water vapor permeability of the films. The antioxidant activity of the films increased with increasing concentrations of the curcumin extract. This study shows the potential benefits of incorporating curcumin extract into chitosan films used as active packaging.

Pectin recovered from mango peel biomass can be used as a potential source for pectic oligosaccharide hydrolysate with excellent probiotic growth-enhancing performance and prebiotic potentials. Consequently, the objectives of the current study were to optimise the enzyme hydrolysis treatment of mango peel pectin (MPP) and to evaluate the pectic oligosaccharide effects of Lactobacillus reuteri DSM 17938 and Bifidobacterium animalis TISTR 2195. Mango of "chok anan" variety was chosen due to its excessive volume of biomass in processing and high pectin content. The optimal treatment for mango peel pectic oligosaccharide (MPOS) valorisation was 24 h of fermentation with 0.3% ( v/v ) pectinase. This condition provided small oligosaccharides with the molecular weight of 643 Da that demonstrated the highest score of prebiotic activity for both of B. animalis TISTR 2195 (7.76) and L. reuteri DSM 17938 (6.87). The major sugar compositions of the oligosaccharide were fructose (24.41% ( w/w )) and glucose (19.52% ( w/w )). For the simulation of prebiotic fermentation, B. animalis TISTR 2195 showed higher proliferation in 4% ( w/v ) of MPOS supplemented (8.92 log CFU/mL) than that of L. reuteri (8.53 CFU/mL) at 72 h of the fermentation time. The main short chain fatty acids (SCFAs) derived from MPOS were acetic acid and propionic acid. The highest value of total SCFA was achieved from the 4% ( w/v ) MPOS supplementation for both of B. animalis (68.57 mM) and L. reuteri (69.15 mM). The result of this study therefore conclusively advises that MPOS is a novel pectic oligosaccharide resource providing the opportunity for the sustainable development approach through utilising by-products from the fruit industry.

Nata de coco has been used as a raw material for food preparation. In this study, the production of carboxymethyl cellulose (CMC) film from nata de coco and the effect of monochloroacetic acid on carboxymethyl bacterial cellulose (CMC n ) and its film were investigated. Bacterial cellulose from nata de coco was modified into CMC form via carboxymethylation using various concentrations of monochloroacetic acid (MCA) at 6, 12, 18, and 24 g per 15 g of cellulose. The results showed that different concentrations of MCA affected the degree of substitution (DS), chemical structure, viscosity, color, crystallinity, and morphology of CMC n . The optimum treatment for carboxymethylation was found using 24 g of MCA per 15 g of cellulose, which provided the highest DS at 0.83. The morphology of CMC n was related to DS value; a higher DS value showed denser and smoother surface than nata de coco cellulose. The various MCA concentrations increased the mechanical properties (tensile strength and percentage of elongation at break) and water vapor permeability of CMC n , which were related to the DS value.

Bacterial cellulose from nata de coco was prepared from the fermentation of coconut juice with Acetobacter xylinum for 10 days at room temperature under sterile conditions. Carboxymethyl cellulose (CMC) was transformed from the bacterial cellulose from the nata de coco by carboxymethylation using different concentrations of sodium hydroxide (NaOH) and monochloroacetic acid (MCA) in an isopropyl (IPA) medium. The effects of various NaOH concentrations on the degree of substitution (DS), chemical structure, viscosity, color, crystallinity, morphology and the thermal properties of carboxymethyl bacterial cellulose powder from nata de coco (CMCn) were evaluated. In the carboxymethylation process, the optimal condition resulted from NaOH amount of 30 g/100 mL, as this provided the highest DS value (0.92). The crystallinity of CMCn declined after synthesis but seemed to be the same in each condition. The mechanical properties (tensile strength and percentage of elongation at break), water vapor permeability (WVP) and morphology of CMCn films obtained from CMCn synthesis using different NaOH concentrations were investigated. The tensile strength of CMCn film synthesized with a NaOH concentration of 30 g/100 mL increased, however it declined when the amount of NaOH concentration was too high. This result correlated with the DS value. The highest percent elongation at break was obtained from CMCn films synthesized with 50 g/100 mL NaOH, whereas the elongation at break decreased when NaOH concentration increased to 60 g/100 mL.

Cellulose from Asparagus officinalis stalk end was extracted and synthesized to carboxymethyl cellulose (CMC as ) using monochloroacetic acid (MCA) via carboxymethylation reaction with various sodium hydroxide (NaOH) concentrations starting from 20% to 60%. The cellulose and CMC as were characterized by the physical properties, Fourier Transform Infrared spectroscopy (FTIR), Differential scanning calorimetry (DSC), Scanning electron microscopy (SEM) and X-ray diffraction (XRD). In addition, mechanical properties of CMC as films were also investigated. The optimum condition for producing CMC as was found to be 30% of NaOH concentration for the carboxymethylation reaction, which provided the highest percent yield of CMC as at 44.04% with the highest degree of substitution (DS) at 0.98. The melting point of CMC as decreased with increasing NaOH concentrations. Crystallinity of CMC as was significantly deformed ( p < 0.05) after synthesis at a high concentration. The L* value of the CMC as was significantly lower at a high NaOH concentration compared to the cellulose. The highest tensile strength (44.59 MPa) was found in CMC as film synthesized with 40% of NaOH concentration and the highest percent elongation at break (24.99%) was obtained in CMC as film treated with 30% of NaOH concentration. The applications of asparagus stalk end are as biomaterials in drug delivery system, tissue engineering, coating, and food packaging.

Thermoplastic elastomer (TPE) was developed by blending thermoplastic starch (TPS) with rubber. Thermoplastic starch-chitosan (TPSC) was prepared by the solution mixing of cassava starch, chitosan (CTS) and glycerol in acidified water (lactic acid 1 wt%) at 80 °C follow by melt mixing at 130 °C. Sodium benzoate (BEN) and chlorhexidine gluconate (Cl) were added during the solution mixing as additives for antimicrobial properties. TPSC was melt-mixed with epoxidized natural rubber (ENR) (70/30 wt/wt). The tensile strength and elongation at break of the TPSC/ENR increased with the additive content. Elastic recovery was improved by the addition of Cl. A new peak in the FTIR data confirmed the reaction between the reactive functional groups of the CTS and the additives with the epoxy groups of ENR. These reactions and miscibility of the TPSC/ENR/additives blends improved the mechanical properties, elasticity, morphology, and antimicrobial properties of the blends., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The aim of this work was to synthesize carboxymethyl cellulose (CMC) and produce CMC films from the cellulose of palm bunch and bagasse agricultural waste. The effect of various amounts of H 2 O 2 (0-40% v / v ) during delignification on the properties of cellulose, CMC, and CMC films was studied. As the H 2 O 2 content increased, yield and the lignin content of the cellulose from palm bunch and bagasse decreased, whereas lightness ( L* ) and whiteness index (WI) increased. FTIR confirmed the substitution of a carboxymethyl group on the cellulose structure. A higher degree of substitution of CMC from both sources was found when 20%-30% H 2 O 2 was employed. The trend in the L* and WI values of each CMC and CMC film was related to those values in their respective cellulose. Bleaching each cellulose with 20% H 2 O 2 provided the cellulose with the highest viscosity and the CMC films with the greatest mechanical (higher tensile strength and elongation at break) and soluble attributes, but the lowest water vapor barrier. This evidence indicates that cellulose delignification with H 2 O 2 has a strong effect on the appearance and physical properties of both CMCs.

This research aimed to synthesize carboxymethyl chitosan (CMCH) from different molecular weights of chitosan including low MW (L, 50-190 kDa), medium MW (M, 210-300 kDa) and high MW (H, 310-375 kDa) on the antioxidant and moisturizing properties. The L-CMCH, M-CMCH and H-CMCH improved the water solubility by about 96%, 90% and 89%, respectively when compared to native chitosan. Higher MW resulted in more viscous of CMCH. For antioxidant properties, IC 50 values of DPPH and ABTS radical scavenging activity for L-CMCH were 1.70 and 1.37 mg/mL, respectively. The L-CMCH had higher antioxidant properties by DPPH and ABTS radical scavenging assay and FRAP. The moisturizing properties on pig skin using a Corneometer ® showed that 0.5% H-CMCH significantly presented ( p ≤ 0.05) greater moisturizing effect than that of untreated-skin, distilled water, propylene glycol and pure chitosan from three molecular weights., Competing Interests: The authors declare no conflict of interest.

Thermoplastic starch (TPS) was prepared by melt-mixing cassava starch with glycerol. Polyethylene (PE), polyethylene-grafted-maleic anhydride (PE-MAH) and poly(lactic acid) (PLA) solutions at 2% ( w/v ) were used to coat TPS using the dip coating process. The tensile strength of TPS increased with the dip coating solution technique, especially for PLA coating. Swelling index, water-soluble matter and water droplet contact angle confirmed the water resistant improvement of TPS by PE-MAH and the PLA dip coating solution. Plasticizer bleeding was found in uncoated TPS after storage, but not in the coated TPS. Coating TPS with PE-MAH and PLA improved the tensile properties, water resistance and conquered plasticizer bleeding problems in TPS.

Corn starch was melt-blended with glycerol (70/30) to prepare thermoplastic starch (TPS) at 140 °C for 10 min. The TPS was melt-blended with polyethylene grafted with maleic anhydride (PE-MAH) and carboxymethyl cellulose (CMC) to improve the properties of the TPS. Phase determination analysis and SEM images demonstrated that the morphology of the TPS/PE-MAH/CMC blend consisted of PE-MAH particles dispersed in the TPS matrix. The tensile strength and elongation were found to have improved with the addition of CMC 5 wt%. Adhesion between the polymers was observed by the asymmetric double-cantilever beam (ADCB) method. The PE-MAH/(TPS + CMC) sample showed high fracture toughness (Gc), which is related to the adhesion energy between the two polymers. This is suggestive of interactions between the MAH groups of PE-MAH and CMC. FT-IR results indicated that the neutralization and aggregation were formed by Na + ions of CMC and the MAH of PE-MAH, which increased the interfacial adhesion between TPS and PE-MAH blend. The improvement in the mechanical properties of the TPS/PE-MAH/CMC blend is concluded to be because of the increase in the interfacial adhesion between TPS and PE-MAH by these neutralization and aggregation processes., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Non-model yeasts within basidiomycetes have considerable importance in agriculture, industry, and environment, but they are not as well studied as ascomycetous yeasts. Serving as a basic technique, nuclear DNA staining is widely used in physiology, ecology, cell biology, and genetics. However, it is unclear whether the classical nuclear DNA staining method for ascomycetous yeasts is applicable to basidiomycetous yeasts. In this study, 5 yeasts ineffectively stained by the classical propidium iodide (PI) staining method were identified from 23 representative basidiomycetous yeasts. Pretreatment of cells using dimethyl sulfoxide (DMSO) or snailase markedly improved cell penetration to PI and thus enabled DNA content determination by flow cytometry on the recalcitrant yeasts. The pretreatments are efficient, simple, and fast, avoiding tedious mutagenesis or genetic engineering used in previous reports. The heterogeneity of cell penetration to PI among basidiomycetous yeasts was attributed to the discrepancy in cell wall polysaccharides instead of capsule or plasma membrane. This study also indicated that care must be taken in attributing PI-negative staining as viable cells when studying non-model microorganisms.

An integrated process for xylooligosaccharides (XOs) and bioethanol production from corncob was investigated. XOs were produced by a consecutive process of KOH treatment and hydrolysis by an in-house thermostable endo-xylanase from Streptomyces thermovulgaris. XO yields of 0.15 g/g KOH-treated corncob (22.13 g/L) and 0.52 g/g raw corncob of cellulose-rich corncob (CRC) were obtained. After 96 h of enzymatic hydrolysis, CRC hydrolysate contained 62.16, 51.21, 10.03 and 0.92 g/L of total sugar, glucose, xylose and arabinose, respectively. Bioethanol production by separate hydrolysis and fermentation (SHF) using CRC hydrolysate, and by simultaneous saccharification and fermentation (SSF) using CRC was studied at 40 °C for thermotolerant Candida glabrata. SHF showed an ethanol yield of 0.28 g/g CRC (21.92 g/L) and ethanol productivity of 0.304 g/L/h with 93% theoretical yield. Surprisingly, by SSF, those parameters were 0.27 g/g CRC (31.32 g/L), 0.33 g/L/h and 89%, respectively. This integrated process might be a new cost-effective approach for corncob valorization., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Saccharification of lignocellulose is a necessary procedure for deconstructing the complex structure for building a sugar platform that can be used for producing biofuel and high-value chemicals. In this study, a carbon-based solid acid catalyst derived from sodium lignosulfonate, a waste by-product from the paper industry, was successfully prepared and used for the hydrolysis of hemicellulose in corncob. The optimum preparation conditions for the catalyst were determined to be carbonization at 250 °C for 6 h, followed by sulfonation with concentrated H 2 SO 4 (98%) and oxidation with 10% H 2 O 2 (solid-liquid ratio of 1 : 75 g mL -1 ) at 50 °C for 90 min. SEM, XRD, FT-IR, elemental analysis and acid-base titration were used for the characterization of the catalysts. It was found that 0.68 mmol g -1 SO 3 H and 4.78 mmol g -1 total acid were loaded onto the catalyst. When corncob was hydrolyzed by this catalyst at 130 °C for 12 h, the catalyst exhibited high selectivity and produced a relatively high xylose yield of up to 84.2% (w/w) with a few by-products. Under these conditions, the retention rate of cellulose was 82.5%, and the selectivity reached 86.75%. After 5 cycles of reuse, the catalyst still showed high catalytic activity, with slightly decreased yields of xylose from 84.2% to 70.7%., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Objectives: A bacterial halotolerant enzyme was characterized to understand the molecular mechanism of salt adaptation and to explore its protein engineering potential., Results: Halotolerant serine protease (Apr&#95;No16) from a newly isolated Bacillus subtilis strain no. 16 was characterized. Multiple alignments with previously reported non-halotolerant proteases, including subtilisin Carlsberg, indicated that Apr&#95;No16 has eight acidic or polar amino acid residues that are replaced by nonpolar amino acids in non-halotolerant proteases. Those residues were hypothesized to be one of the primary contributors to salt adaptation. An eightfold mutant substituted with Ala residues exhibited 1.2- and 1.8-fold greater halotolerance at 12.5% (w/v) NaCl than Apr&#95;No16 and Carlsberg, respectively. Amino acid substitution notably shifted the theoretical pI of the eightfold mutant, from 6.33 to 9.23, compared with Apr&#95;No16. The resulting protein better tolerated high salt conditions., Conclusions: Changing the pI of a bacterial serine protease may be an effective strategy to improve the enzyme's halotolerance.

Candida shehatae ATCC 22984, a xylose-fermenting yeast, showed an ability to produce ethanol in both glucose and xylose medium. Maximum ethanol produced by the yeast was 48.8 g/L in xylose and 52.6 g/L in glucose medium with ethanol yields that varied between 0.3 and 0.4 g/g depended on initial sugar concentrations. Xylitol was a coproduct of ethanol production using xylose as substrate, and glycerol was detected in both glucose and xylose media. Kinetic model equations indicated that growth, substrate consumption, and product formation of C. shehatae were governed by substrate limitation and inhibition by ethanol. The model suggested that cell growth was totally inhibited at 40 g/L of ethanol and ethanol production capacity of the yeast was 52 g/L, which were in good agreement with experimental results. The developed model could be used to explain C. shehatae fermentation in glucose and xylose media from 20 to 170 g/L sugar concentrations.

A lactic acid producing bacterium, Lactobacillus rhamnosus M-23, newly isolated from a rice washing drainage storage tank was found to produce l-(+)-lactic acid from a non-sterilized mixture of rice washing drainage and rice bran without any additions of nutrients under the simultaneous saccharification and fermentation (SSF) process. This strain has the ability to utilize the non-sterilized rice washing drainage and rice bran as a source of carbohydrate, saccharifying enzymes and nutrients for lactic acid production. Observation of extracellular protease activity in SSF culture broth showed that a higher protease activity was present in strain M-23 than in other isolated lactic acid producing bacteria (LABs). To investigate the structural changes of solid particles of rice washing drainage throughout LAB cultivation, scanning electron microscopic (SEM) observation and Fourier transform infrared-spectroscopy (FT-IR) analysis were performed. The results of the SEM observation showed that the surface material could be removed from solid particles of rice washing drainage treated by culture broth (supernatant) of strain M-23, thus exposing the crystal structure of the starch particle surface. The results of the FT-IR analysis revealed that the specific transmittance decrease of the CC and CO stretching and OH group of the solid particles of the rice washing drainage were highly correlated with the produced lactic acid concentration and extracellular protease activity, respectively. These results demonstrate the high lactic acid producing ability of strain M-23 from a non-sterilized mixture of rice washing drainage and rice bran under the SSF condition due to the removal of proteinaceous material and exposure of the starch particle surface by extracellular protease., (Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Cassava starch was melt-blended with glycerol (70/30wt%/wt%) at 140°C to prepare thermoplastic starch (TPS). Chitosan (CTS) was premixed with starch and glycerol, in acidified water (lactic acid 2wt%), at 1, 5 and 10wt%/wt%. TPS/CTS was then melt-blended (160°C) with polyethylene-graft-maleic anhydride (PE-MAH). Phase determination and scanning electron microscopy indicated TPS/PE-MAH/CTS had a co-continuous morphology and CTS-induced phase inversion to give dispersed PE-MAH particles in a TPS matrix. Tensile strength at break and elongation, melt viscosity, fracture toughness and water contact angle of TPS/PE-MAH were improved by CTS incorporation. TPS/PE-MAH/CTS blends decreased the melting temperature of TPS and PE-MAH compared to the neat polymers. FTIR confirmed a reaction had occurred between amino groups (NH2) of CTS and the MAH groups of PE-MAH. This reaction and the enhanced miscibility between TPS and CTS improved the mechanical properties of the TPS/PE-MAH/CTS blend, particularly at 5wt%/wt% CTS., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study aimed to investigate the efficiency of an aerobic sequencing batch reactor (aerobic SBR) in a nonsterile system using the application of an experimental design via central composite design (CCD). The acidic whey obtained from lactic acid fermentation by immobilized Lactobacillus plantarum sp. TISTR 2265 was fed into the bioreactor of the aerobic SBR in an appropriate ratio between acidic whey and cheese whey to produce an acidic environment below 4.5 and then was used to support the growth of Dioszegia sp. TISTR 5792 by inhibiting bacterial contamination. At the optimal condition for a high yield of biomass production, the system was run with a hydraulic retention time (HRT) of 4 days, a solid retention time (SRT) of 8.22 days, and an acidic whey concentration of 80% feeding. The chemical oxygen demand (COD) decreased from 25,230 mg/L to 6,928 mg/L, which represented a COD removal of 72.15%. The yield of biomass production and lactose utilization by Dioszegia sp. TISTR 5792 were 13.14 g/L and 33.36%, respectively, with a long run of up to 180 cycles and the pH values of effluent were rose up to 8.32 without any pH adjustment.

Cheese whey contains a high organic content and causes serious problems if it is released into the environment when untreated. This study aimed to investigate the optimum condition of lactic acid production using the microaerobic sequencing batch reactor (microaerobic SBR) in a nonsterile system. The high production of lactic acid was achieved by immobilized Lactobacillus plantarum TISTR 2265 to generate an acidic pH condition below 4.5 and then to support single-cell protein (SCP) production in the second aerobic sequencing batch reactor (aerobic SBR). A hydraulic retention time (HRT) of 4 days and a whey concentration of 80% feeding gave a high lactic acid yield of 12.58 g/L, chemical oxygen demand (COD) removal of 62.38%, and lactose utilization of 61.54%. The microbial communities in the nonsterile system were dominated by members of lactic acid bacteria, and it was shown that the inoculum remained in the system up to 330 days.

Two amylases, amylase I and amylase II from Bacillus subtilis strain FP-133, were purified to homogeneity and characterized. Their stabilities toward temperature, pH, and organic solvents, and their substrate specificities toward polysaccharides and oligosaccharides were similar. Under moderately high salt conditions, both amylases were more stable than commercial B. licheniformis amylase, and amylase I retained higher amylase activity than amylase II. The N-terminal amino acid sequence, genomic southern blot analysis, and MALDI-TOFF-MS analysis indicated that the halotolerant amylase I was produced by limited carboxy-terminal truncation of the amylase II peptide. The deduced amino acid sequence of amylase II was >95% identical to that of previously reported B. subtilis α-amylases, but their carboxy-terminal truncation points differed. Three recombinant amylases--full-length amylase corresponding to amylase II, an artificially truncated amylase corresponding to amylase I, and an amylase with a larger artificial C-terminal truncation--were expressed in B. subtilis. The artificially truncated recombinant amylases had the same high amylase activity as amylase I under moderately high salt conditions. Sequence comparisons indicated that an increased ratio of Asp/Glu residues in the enzyme may be one factor responsible for increasing halotolerance., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The optimal culture condition for extracellular recombinant single chain variable fragment anti HIV-1 p17 protein (scFv anti-p17) production in Escherichia coli HB2151 was investigated by the sequential simplex optimization (SS) method. Five variable parameters were submitted in the fermentation process. The most favorable condition obtained from 19 independent experiments was as followed: 58 µM of IPTG induction to 1.7 OD600 nm at 25.5°C for 16 h with 202 rpm agitation rate. The amount of secreted scFv anti-p17 at the optimal condition was 38% higher than under the control condition. The binding activity of soluble extracellular scFv anti-p17 protein increased 95.5% and 73.2% in comparison with the control condition and non-optimized condition respectively. The soluble scFv anti-p17 from crude HB2151 lysated was subsequently purified by immobilized metal ion affinity chromatography (IMAC) with His-tag. The purified scFv anti-p17 was intact and retained its antigen-binding affinity against HIV-1 p17. We demonstrated that the sequential simplex optimization method was a key for exertion of high yield with fewer experimental requirements for acquiring of large scale secretory protein production.

It is believed that high concentrations of sodium chloride (NaCl) suppress the biosynthesis of exopolysaccharide (EPS) in lactic acid bacteria (LAB). Nevertheless, overproduction of EPSs due to high salinity stress in solid state fermentation performed on an agar surface was demonstrated in this study using a response surface methodology via a central composite design (CCD). Under optimized conditions with NaCl 4.97% and sucrose 136.5 g/L at 40.79 h of incubation, the EPS yield was 259% (86.36 g/L of EPS), higher than the maximum yield produced with the modified MRS medium containing only 120 g/L of sucrose without NaCl (33.4 g/L of EPS). Biosynthesis of EPS by Lactobacillus confusus TISTR 1498 was independent of biomass production. Our results indicated that high salinity stress can enhance EPS production in solid state fermentation.

Cellulase-free xylanase production by thermophilic Streptomyces thermovulgaris TISTR1948 was cultivated in a basal medium with rice straw as sole source of carbon and as an inducible substrate. Variable medium components were selected in accordance with the Plackett-Burman experimental design. The optimization conditions of physical factors (pH and temperature levels) were then combined in further studies through the response surface methodology approach. Only two significant components, rice straw and yeast extract, were chosen for the optimization studies. A second-order quadratic model was constructed by central composite design (CCD). The model revealed that both pH and temperature levels were significant, and were dependent on xylanase production. Under these experimental designs, the xylanase yield increased from 51.11 to 274.49 U/mL (3,400 to 10,000 U/g of rice straw) or about 537% higher than an unoptimized basal medium. The optimum conditions to achieve maximum yield of xylanase were 27.45 g/L of rice straw and 5.42 g/L of yeast extract under relatively neutral conditions of pH 7.11, 50.03 °C, and a incubation period.

Initial rate and biotransformation studies were applied to refine and validate a mathematical model for enzymatic (R)-phenylacetylcarbinol (PAC) production from pyruvate and benzaldehyde using Candida utilis pyruvate decarboxylase (PDC). The rate of PAC formation was directly proportional to the enzyme activity level up to 5.0 U ml-1 carboligase. Michaelis-Menten kinetics were determined for the effect of pyruvate concentration on the reaction rate. The effect of benzaldehyde followed the sigmoidal shape of the Monod-Wyman-Changeux (MWC) model. The biotransformation model, which also included a term for PDC inactivation by benzaldehyde, was used to determine the overall rate constants for the formation of PAC, acetaldehyde, and acetoin. These values were determined from data for three batch biotransformations performed over a range of initial concentrations (viz. 50-150 mM benzaldehyde, 60-180 mM pyruvate, 1.1-3.4 U ml-1 enzyme activity). The finalized model was then used to predict a batch biotransformation profile at 120/100 mM initial pyruvate/benzaldehyde (initial enzyme activity 3.0 U ml-1). The simulated kinetics gave acceptable fitting (R2 = 0.9963) to the time courses of these latter experimental data for substrates pyruvate and benzaldehyde, product PAC, by-products acetaldehyde and acetoin, as well as enzyme activity level.