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Benchmarking hydrolytic potential of cellulase cocktail obtained from mutant strain of Talaromyces verruculosus IIPC 324 with commercial biofuel enzymes.

ABSTRACT: In the present study, an attempt was made to benchmark the hydrolytic potential of cellulase cocktail obtained from stable mutant UV-8 of Talaromyces verruculosus IIPC 324 (NFCCI 4117) with three commercially available cellulases. With two experimental approaches, acid-pretreated sugarcane bagasse was subjected to hydrolysis for 72 h, where all the enzymes were dosed on the basis of common protein or common cellulase activity /g cellulose content. Concentrated fungal enzyme (CFE) of mutant UV-8 resulted in ~ 59% and 55% saccharification of acid-pretreated sugarcane bagasse after 72 h at 55 °C and pH 4.5 with respect to reducing sugar release, when dosed at 25 mg protein/g and 500 IU CMC'ase/g cellulose, respectively. On the other hand, at similar dosages, the performance of Cellic CTec2 was best resulting in 77% and 66% saccharification, respectively. When enzyme desorption studies were undertaken by carrying out cellulase activities in saccharified broth after 72 h CFE of UV-8 emerged as the best cellulase cocktail. A minimum of 90% endoglucanase and 60% cellobiohydrolase I was successfully desorbed from residual biomass, thereby increasing the probability of enzyme recycle and reuse for next round of hydrolysis.

SUBMITTER: Jain L

PROVIDER: S-EPMC6314943 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Benchmarking hydrolytic potential of cellulase cocktail obtained from mutant strain of <i>Talaromyces verruculosus</i> IIPC 324 with commercial biofuel enzymes.

Jain Lavika L Kurmi Akhilesh Kumar AK Agrawal Deepti D

3 Biotech 20190102 1

In the present study, an attempt was made to benchmark the hydrolytic potential of cellulase cocktail obtained from stable mutant UV-8 of <i>Talaromyces verruculosus</i> IIPC 324 (NFCCI 4117) with three commercially available cellulases. With two experimental approaches, acid-pretreated sugarcane bagasse was subjected to hydrolysis for 72 h, where all the enzymes were dosed on the basis of common protein or common cellulase activity /g cellulose content. Concentrated fungal enzyme (CFE) of mutan ...[more]

PMID: 30622861

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Project description:BACKGROUND:A complete saccharification of plant polymers is the critical step in the efficient production of bio-alcohols. Beta-glucosidases acting in the degradation of intermediate gluco-oligosaccharides produced by cellulases limit the yield of the final product. RESULTS:In the present work, we have identified and then successfully cloned, expressed, purified and characterised 4 highly active beta-glucosidases from fibre-adherent microbial community from the cow rumen. The enzymes were most active at temperatures 45-55°C and pH 4.0-7.0 and exhibited high affinity and activity towards synthetic substrates such as p-nitrophenyl-beta-D-glucopyranoside (pNPbetaG) and pNP-beta-cellobiose, as well as to natural cello-oligosaccharides ranging from cellobiose to cellopentaose. The apparent capability of the most active beta-glucosidase, herein named LAB25g2, was tested for its ability to improve, at low dosage (31.25 units g-1 dry biomass, using pNPbetaG as substrate), the hydrolysis of pre-treated corn stover (dry matter content of 20%; 350 g glucan kg-1 dry biomass) in combination with a beta-glucosidase-deficient commercial Trichoderma reseei cellulase cocktail (5 units g-1 dry biomass in the basis of pNPbetaG). LAB25g2 increased the final hydrolysis yield by a factor of 20% (44.5 ± 1.7% vs. 34.5 ± 1.5% in control conditions) after 96-120 h as compared to control reactions in its absence or in the presence of other commercial beta-glucosidase preparations. The high stability (half-life higher than 5 days at 50°C and pH 5.2) and 2-38000 fold higher (as compared with reported beta-glucosidases) activity towards cello-oligosaccharides may account for its performance in supplementation assays. CONCLUSIONS:The results suggest that beta-glucosidases from yet uncultured bacteria from animal digestomes may be of a potential interest for biotechnological processes related to the effective bio-ethanol production in combination with low dosage of commercial cellulases.

Project description:BackgroundThe discovery of enzymes named lytic polysaccharide monooxygenases (LPMOs) has had a major impact on the efficiency of current commercial cellulase cocktails for saccharification of lignocellulosic biomass. However, the notion that LPMOs use molecular oxygen as a co-substrate and require two externally delivered electrons per catalytic cycle poses a challenge in the development of efficient large-scale industrial processes. Building on the recent discovery that H2O2, rather than O2, is the co-substrate of LPMOs, we show here how cellulose degradation by the LPMO-containing commercial cellulase cocktail Cellic® CTec2 can be controlled and boosted by supplying the reaction with H2O2.ResultsThe controlled supply of anaerobic hydrolysis reactions with H2O2 and sub-stoichiometric amounts of reductant increased apparent LPMO activity by almost two orders of magnitude compared to standard aerobic reactions utilizing O2 and stoichiometric amounts of reductant. Improved LPMO activity was correlated with enhanced saccharification rates and yields for a model cellulosic substrate (Avicel) as well as industrial lignocellulosic substrates (sulfite-pulped Norway spruce and steam-exploded birch), although the magnitude of the effects was substrate dependent. Improvements in lignocellulose conversions were achieved at low H2O2 feeding rates (in the range of 90-600 µM h-1). Tight control of LPMO reactions by controlled supply of H2O2 under anaerobic conditions was possible.ConclusionWe report saccharification rates and yields for a model substrate (Avicel) and industrial lignocellulosic substrates that, at low H2O2 feeding rates, are higher than those seen under standard aerobic conditions. In an industrial setting, controlling and supplying molecular oxygen and stoichiometric amounts of reductant are challenging. The present report shows that the use of small amounts of a liquid bulk chemical, H2O2, provides an alternative to the currently available processes, which likely is cheaper and more easy to control, while giving higher product yields.

Project description:ImportanceCommercial healthcare claims datasets represent a sample of the US population that is biased along socioeconomic/demographic lines; depending on the target population of interest, results derived from these datasets may not generalize. Rigorous comparisons of claims-derived results to ground-truth data that quantify this bias are lacking.Objectives(1) To quantify the extent and variation of the bias associated with commercial healthcare claims data with respect to different target populations; (2) To evaluate how socioeconomic/demographic factors may explain the magnitude of the bias.DesignThis is a retrospective observational study. Healthcare claims data come from the Merative™ MarketScan® Commercial Database; reference data for comparison come from the State Inpatient Databases (SID) and the US Census. We considered three target populations, aged 18-64 years: (1) all Americans; (2) Americans with health insurance; (3) Americans with commercial health insurance.ParticipantsWe analyzed inpatient discharge records of patients aged 18-64 years, occurring between 01/01/2019 to 12/31/2019 in five states: California, Iowa, Maryland, Massachusetts, and New Jersey.OutcomesWe estimated rates of the 250 most common inpatient procedures, using claims data and using reference data for each target population, and we compared the two estimates.ResultsThe average rate of inpatient discharges per 100 person-years was 5.39 in the claims data (95% CI: [5.37, 5.40]) and 7.003 (95% CI: [7.002, 7.004]) in the reference data for all Americans, corresponding to a 23.1% underestimate from claims. We found large variation in the extent of relative bias across inpatient procedures, including 22.8% of procedures that were underestimated by more than a factor of 2. There was a significant relationship between socioeconomic/demographic factors and the magnitude of bias: procedures that disproportionately occur in disadvantaged neighborhoods were more underestimated in claims data (R 2 51.6%, p < 0.001). When the target population was restricted to commercially insured Americans, the bias decreased substantially (3.2% of procedures were biased by more than factor of 2), but some variation across procedures remained.Conclusions and relevanceNaïve use of healthcare claims data to derive estimates for the underlying US population can be severely biased. The extent of bias is at least partially explained by neighborhood-level socioeconomic factors.

Dataset Information

Benchmarking hydrolytic potential of cellulase cocktail obtained from mutant strain of Talaromyces verruculosus IIPC 324 with commercial biofuel enzymes.

Publications

Benchmarking hydrolytic potential of cellulase cocktail obtained from mutant strain of <i>Talaromyces verruculosus</i> IIPC 324 with commercial biofuel enzymes.

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