Thanongsak Chaiyaso

Optimization of cellulase-free xylanase production by thermophilic streptomyces thermovulgaris TISTR1948 through Plackett-Burman and response surface methodological approaches

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.

Eco-friendly processing in enzymatic xylooligosaccharides production from corncob: Influence of pretreatment with sonocatalytic–synergistic Fenton reaction and its antioxidant potentials

Delignification can be considered as a feasible process to pretreat lignocellulosic biomass in xylooligosaccharides production after the performance and efficiency has been improved through a few modifications. This study compared various pretreatment strategies such as Fenton, sonocatalytic, and sonocatalytic-synergistic Fenton employed on corncob in order to expose lignin content and saccharides to enhance the xylooligosaccharides yield by enzymatic hydrolysis. The dissolution of lignin and xylooligosaccharides production of corncob was enhanced by ultrasound assisted TiO2 and Fenton reaction. The corncob pretreated with a sonocatalytic-synergistic Fenton reaction gave the highest release of the lignin concentration level (1.03 g/L), dissolution level (80.25%), and xylooligosaccharides content (46.45 mg/g substrate). A two-step pretreatment processes consisting of the alkali treatment (pretreatment) and sonocatalytic-synergistic Fenton process (posttreatment) illustrated that subsequent enzymatic hydrolysis could be enhanced considerably. The release of the lignin concentration and xylooligosaccharides content were 33.20 g/L and 174.81 mg/g substrate, respectively. The antioxidant potential of xylooligosaccharides showed significant differences regarding the amount of xylooligosaccharides and the phenolic compounds produced.

Production of xylooligosaccharides from corncob using a crude thermostable endo-xylanase from Streptomyces thermovulgaris TISTR1948 and prebiotic properties

Production of xylooligosaccharides (XOs) from corncob using the thermostable endo-xylanase from Streptomyces thermovulgaris TISTR1948 was investigated using KOH pretreatment, followed by enzymatic hydrolysis. The optimal reaction time for production of XOs was 12 h, after which xylobiose comprised a majority of products, and a low xylose content was observed. The optimal conditions for production of XOs were studied using a central composite design. At an enzyme concentration of 129.43 U/g of substrate, 53.80°C, and pH 6.17, the yield of XOs reached 162.97 mg/g of substrate or 752.15 mg/g of hemicellulose in KOH-pretreated corncob. The prebiotic properties of XOs derived from corncob were also investigated using in vitro fermentation of those XOs with the known probiotic strains Lactobacillus casei TISTR1463, L. lactis TISTR1464, and L. plantarum TISTR1465. XOs derived from corncob were comparable to commercial XOs for an ability to enhance the growth of the specified probiotic lactobacilli.

Optimization of Exopolysaccharide Overproduction by Lactobacillus confusus in Solid State Fermentation under High Salinity Stress

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.

Characterization of the native form and the carboxy‐terminally truncated halotolerant form of α‐amylases from Bacillus subtilis strain FP‐133

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.

Green and chemical-free process of enzymatic xylooligosaccharide production from corncob: Enhancement of the yields using a strategy of lignocellulosic destructuration by ultra-high pressure pretreatment

In this study, the pressures at 50-500MPa were evaluated at different time to pretreat and further enzyme hydrolysis. The ultra-high pressure (UHP) pretreatment at 100MPa for 10min led to improved accessibility of enzyme for conversion of xylan to xylooligosaccharide (XOS). The maximum XOS yield of 35.6mg/g substrate was achieved and firstly reported at 10% (w/v) of substrate, 100U of endo-xylanase/g corncobs and incubation time of 18h. The enzymatic hydrolysis efficiency was increased by 180.3% and released a high amount of xylobiose. The UHP pretreatment relatively did not affect to the composition of corncob, but decreased 34.3% of lignin. Interestingly, antioxidant activities of XOS using UHP pretreatment were higher than untreated corncob. The UHP pretreatment improved lignocellulosic destructuration and XOS yields in a shorter time without the need of chemicals, implying that UHP could be an effective pretreatment of biomass with a chemical-free process.

Influence of food colorant and initial COD concentration on the efficiencies of micro-aerobic sequencing batch reactor (micro-aerobic SBR) for casein recovery under non-sterile condition by Lactobacillus casei TISTR 1500.

The acid biocoagulants produced from non-sterile lactic acid fermentation by Lactobacillus casei TISTR 1500 were used to settle colloidal protein, mainly casein, at the isoelectric point in dairy effluent prior to secondary treatment. High concentration of azo dye (Ponceau 4R) in the dairy wastewater and the stress of starvation decreased the efficiencies of the micro-aerobic SBR. Consequently, low casein recovery obtained and organic removal suffered a decline. The number of lactic acid bacteria (LAB) also declined from log 7.4 to log 5.30 in the system fed with 400 mg L(-1) of the dye containing wastewater. The recovery of the system, however, showed that 25,000 mg COD L(-1) influent with 200 mg L(-1) of the dye maintained the growth of LAB in the range of log 7.74-8.12, with lactic and acetic production (2597 and 197 mg L(-1)) and 83% protein removal. The results in this study suggested that the inhibitory effects were compensated with high organic content feeding.

Treatability of cheese whey for single-cell protein production in nonsterile systems: Part II. The application of aerobic sequencing batch reactor (aerobic SBR) to produce high biomass of Dioszegia sp. TISTR 5792

ABSTRACT 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.

Direct bioconversion of rice residue from canteen waste into lipids by new amylolytic oleaginous yeast Sporidiobolus pararoseus KX709872

ABSTRACT The new amylolytic oleaginous red yeast, Sporidiobolus pararoseus KX709872, produced both α-amylase (540 ± 0.09 mU/mL) and amyloglucosidase (23 ± 0.00 mU/mL) and showed good ability to directly convert rice residue from canteen waste to biomass and lipids. Effects of medium composition and cultivation conditions on growth and lipid accumulation for strain KX709872 were investigated under shaking flask and upscaling levels. At C : N ratio of 25 : 1, pH 5.45, 22.36°C, and 199.40 rpm for 7 days, volumetric production of biomass and lipids, lipid content, and lipid productivity reached 17.69 ± 0.44, 8.35 ± 0.19 g/L, 49.48 ± 0.41% (w/w), and 1.67 ± 0.11 g/L/day, respectively. Production of lipids was also implemented in 5.0-L stirred tank bioreactor with 2.5 L of optimized medium at 300 rpm and 3.0 vvm for 5 days. Volumetric production of biomass and lipids, lipid content, and lipid productivity were 16.33 ± 0.49, 8.75 ± 0.13 g/L, 56.61 ± 0.04% (w/w), and 2.19 ± 0.03 g/L/day, respectively. Meanwhile, the fatty acids of lipids from strain KX709872 had high oleic acid content (60−62%) which was similar to those of vegetable oils, indicating that these lipids are promising as an alternative biodiesel feedstock. Moreover, the biodiesel derived from lipids of strain KX709872 had properties satisfying the criteria of ASTM D6751 and EN 14214 standards.

Enhancement of carotenoids and lipids production by oleaginous red yeast sporidiobolus pararoseus KM281507

ABSTRACT Bioconversion of biodiesel-derived crude glycerol into carotenoids and lipids was investigated by a microbial conversion of an oleaginous red yeast Sporidiobolus pararoseus KM281507. The methanol content in crude glycerol (0.5%, w/v) did not show a significant effect on biomass production by strain KM281507. However, demethanolized crude glycerol significantly supported the production of biomass (8.64 ± 0.13 g/L), lipids (2.92 ± 0.03 g/L), β-carotene (15.76 ± 0.85 mg/L), and total carotenoids (33.67 ± 1.28 mg/L). The optimal conditions suggested by central composite design were crude glycerol concentration (55.04 g/L), initial pH of medium (pH 5.63) and cultivation temperature (24.01°C). Under these conditions, the production of biomass, lipids, β-carotene, and total carotenoids were elevated up to 8.83 ± 0.05, 4.00 ± 0.06 g/L, 27.41 ± 0.20, and 53.70 ± 0.48 mg/L, respectively. Moreover, an addition of olive oil (0.5 − 2.0%) dramatically increased the production of biomass (14.47 ± 0.15 g/L), lipids (6.40 ± 0.09 g/L), β-carotene (54.43 ± 0.95 mg/L), and total carotenoids (70.92 ± 0.51 mg/L). The oleic acid content in lipids was also increased to 75.1% (w/w) of total fatty acids, indicating a good potential to be an alternative biodiesel feedstock. Meanwhile, the β-carotene content in total carotenoids was increased to 76.7% (w/w). Hence, strain KM281507 could be a good potential source of renewable biodiesel feedstock and natural carotenoids.