Effect of Tween 80 on Hydrolytic Activity and Substrate Accessibility of Carbohydrolase I (CBH I) from Trichoderma viride

Kim, Wan-Jae; Gamo, Y.; Sani, Yahaya Mohammed; Wusiman, Yimiti; Ogawa, Satoshi; Karita, Shuichi; Goto, Masakazu

doi:10.5713/ajas.2006.684

Cited by 30 publications

(16 citation statements)

References 28 publications

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“…3). Kim et al (2006) reported that the substrate accessibility and hydrolytic activity of CBH I enzyme were increased by addition of tween 80 in T. viride by decreasing the absorption of enzyme on to cellulose. Additionally, supplementation of PEG 6000 to the crude enzyme mixture increased the cellulase activity linearly with the concentration of PEG.…”

Section: Effect Of Additives On Cellulases Productionmentioning

confidence: 99%

Use of combined UV and chemical mutagenesis treatment of Aspergillus terreus D34 for hyper-production of cellulose-degrading enzymes and enzymatic hydrolysis of mild-alkali pretreated rice straw

Kumar

Parikh

Singh

et al. 2015

Bioresour. Bioprocess.

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Background: Microbial production of cellulose-degrading enzymes could be significantly improved using traditional mutagenesis treatment. Development of high-titre cellulase producing mutants drastically reduces the costs involved in cellulase production and downstream processing in commercial-scale enzyme production. Here, we have evaluated the efficacy of different Aspergillus terreus D34 mutants for hyper-production of improved cellulase enzymes utilizing locally available lignocellulosic biomass residues as growth substrates in solid state fermentation conditions. Further, enzymatic hydrolysis of mild-alkali pre-treated rice straw was performed using the improved cellulases.Results: A 4.9-fold higher β-glucosidase activity was obtained from ethyl methyl sulphonate (EMS) treated mutant strain (EMS2) when grown on mixed rice straw/sugarcane bagasse (RSBG) biomass growth substrate. Similarly with the EMS2 mutant and BG-grown culture extract a 1.1-fold higher xylanase activity was observed. Irrespective of the growth substrates and the mutant strains, the maximum cellulase (FPase, carboxymethyl cellulase, avicelase, β-glucosidase) and xylanase activities (U mL −1 ) were 2.34, 39.8, 2.46, 19.9 and 655, respectively. Further, external supplementation of 20% bovine serum albumin (BSA), 3% tween 80 and 20% polyethylene glycol (PEG) 6000 to the crude enzyme extract increased the FPase activity nearly 4.0-, 2.8-and 2.2-fold. Addition of 0.05% sodium benzoate marginally increased the stability of cellulase enzyme and retained more than 60% of the initial activity after 96 h incubation at 37°C. While at 4°C, no loss in enzyme activity was observed even after prolonged incubation period (up to 90 days). Further, maximum reducing sugars of 0.842 g g −1 at a rate of 0.25 mM g −1 h −1 at 10% biomass loading of mild-alkali pretreated rice straw was produced using the BG-grown culture extract of EMS2 mutant strain. Conclusion:The extracellular protein production and corresponding cellulase activities of A. terreus D34 were significantly enhanced after combined UV and chemical mutagenesis treatments. In the present study, besides accelerating the rate of cellulase production, we have also demonstrated production of high reducing sugars by enzymatic saccharification of pretreated lignocellulosic biomass using hyper-produced cellulase enzymes. Due to high enzyme activity of the cellulase enzymes produced from the mutant strains, the volume of enzyme loadings in enzymatic hydrolysis could be reduced up to 7-fold. These studies clearly show the potential of the developed hyper-cellulase producing mutants in decreasing the overall process economics of cellulosic ethanol technology.

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Section: Effect Of Additives On Cellulases Productionmentioning

confidence: 99%

Use of combined UV and chemical mutagenesis treatment of Aspergillus terreus D34 for hyper-production of cellulose-degrading enzymes and enzymatic hydrolysis of mild-alkali pretreated rice straw

Kumar

Parikh

Singh

et al. 2015

Bioresour. Bioprocess.

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“…The mechanism is still not entirely clear, but such additives have been thought to impede deactivation and/or unproductive binding, increase cellulose accessibility, and/or enhance enzyme activity (Eriksson et al, 2002;Huang et al, 2003;Kim et al, 2006;Mizutani et al, 2002;Sewalt et al, 1997;Yang and Wyman, 2006).…”

Section: Introductionmentioning

confidence: 99%

Effect of additives on the digestibility of corn stover solids following pretreatment by leading technologies

Kumar

Wyman²

2009

Biotech & Bioengineering

132

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Bovine serum albumin (BSA), Tween-20, and polyethylene glycol (PEG6000) were added to washed corn stover solids produced by ammonia fiber expansion (AFEX), ammonia recycled percolation (ARP), dilute sulfuric acid (DA), lime, controlled pH, and sulfur dioxide (SO(2)) pretreatments and to untreated corn stover (UT) and pure Avicel glucan prior to adding cellulase supplemented with beta-glucosidase at an activity ratio of 1:2/g and a moderate enzyme loading of 16.1 mg/g glucan in the raw corn stover. The additives were applied individually at 150, 300, and 600 mg/g glucan in the pretreated solids and in combinations of equal amounts of each that totaled 600 mg/g. The greatest increase in total sugar release was by Tween-20 with SO(2) pretreated solids followed by PEG6000 with ARP solids and Tween-20 with lime solids. The effectiveness of the additives was observed to depend on the type of sugars left in the solids, suggesting that it may be more beneficial to use the mixture of these additives to realize a high total sugar yield. In addition, little enhancement in sugar release was possible beyond a loading of 150 mg additives/g glucan for most pretreatments, and combinations did not improve sugar release much over use of additives alone for all except SO(2). Additives were also found to significantly increase concentrations of cellobiose and cellooligomers after 72 h of Avicel hydrolysis.

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“…Additionally, Tween 80 decreases the absorption of enzymes to substrates, which might be functional in maintaining enzymatic reactivity and stability. 39,40) The growth enhancing effect of Tween 80 can increase the nutrient intake of cellulolytic bacterium R. albus 7, and it improved its bacteriocin production in this study. A previous report indicated that phenylpropanoic acid (PPA) accounted for part of the stimulatory effect of rumen fluid on the growth rate of R. albus 8.…”

Section: Discussionmentioning

confidence: 61%

Production and Characterization of a Bacteriocin from Ruminal BacteriumRuminococcus albus7

Wang

Chen

Hsu

2012

Bioscience, Biotechnology, and Biochemistry

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The characteristics of a bacteriocin from Ruminococcus albus 7 and its potential as an antibiotic alternative were examined in this study. The addition of 3 µM 3-phenylpropanoic acid (PPA) and 0.2% Tween 80 to the culturing medium improved bacteriocin production by 2.5-fold. Native polyacrylamide gel electrophoresis of the antagonistically active gel filtration fraction established that the molecular weight of the R. albus 7 bacteriocin was approximately 36 kDa. The bacteriocin was sensitive to pepsin, protease, and pancreatin, and was inactivated by heating at 65 °C for 1 h. Simulating in vitro avian digestion decreased the antagonistic activity by 74.7%, but the addition of 1% bovin serum albumin restored 13% of the lost antagonistic activity. Following ion-exchange purification, the bacteriocin had sufficient antagonistic activity against five tested pathogenic strains, but the addition of a protectant is necessary for utilization of bacteriocin of R. albus 7 as an antibiotic alternative in animal feed.

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Effect of Tween 80 on Hydrolytic Activity and Substrate Accessibility of Carbohydrolase I (CBH I) from Trichoderma viride

Cited by 30 publications

References 28 publications

Use of combined UV and chemical mutagenesis treatment of Aspergillus terreus D34 for hyper-production of cellulose-degrading enzymes and enzymatic hydrolysis of mild-alkali pretreated rice straw

Use of combined UV and chemical mutagenesis treatment of Aspergillus terreus D34 for hyper-production of cellulose-degrading enzymes and enzymatic hydrolysis of mild-alkali pretreated rice straw

Effect of additives on the digestibility of corn stover solids following pretreatment by leading technologies

Production and Characterization of a Bacteriocin from Ruminal BacteriumRuminococcus albus7

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