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Thermostable Cellulase Biosynthesis from Paenibacillus alvei and its Utilization in Lactic Acid Production by Simultaneous Saccharification and Fermentation.

ABSTRACT: Cellulosic date palm wastes may have beneficial biotechnological applications for eco-friendly utilization. This study reports the isolation of thermophilic cellulase-producing bacteria and their application in lactic acid production using date palm leaves. The promising isolate was identified as Paenibacillus alvei by 16S rRNA gene sequencing. Maximum cellulase production was acquired using alkaline treated date palm leaves (ATDPL) at 48 h and yielded 4.50 U.mL^-1 FPase, 8.11 U.mL^-1 CMCase, and 2.74 U.mL^-1 β-glucosidase. The cellulase activity was optimal at pH 5.0 and 50°C with good stability at a wide temperature (40-70°C) and pH (4.0-7.0) range, demonstrating its suitability in simultaneous saccharification and fermentation. Lactic acid fermentation was optimized at 4 days, pH 5.0, 50°C, 6.0% cellulose of ATDPL, 30 FPU/ g cellulose, 1.0 g. L^-1 Tween 80, and 5.0 g. L^-l yeast extract using Lactobacillus delbrueckii. The conversion efficiency of lactic acid from the cellulose of ATDPL was 98.71%, and the lactic acid productivity was 0.719 g. L^-1 h^-1. Alkaline treatment exhibited a valuable effect on the production of cellulases and lactic acid by reducing the lignin content and cellulose crystallinity. The results of this study offer a credible procedure for using date palm leaves for microbial industrial applications.

SUBMITTER: Mostafa YS

PROVIDER: S-EPMC8114780 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundSimultaneous saccharification and fermentation (SSF) of pre-treated lignocellulosics to biofuels and other platform chemicals has long been a promising alternative to separate hydrolysis and fermentation processes. However, the disparity between the optimum conditions (temperature, pH) for fermentation and enzyme hydrolysis leads to execution of the SSF process at sub-optimal conditions, which can affect the rate of hydrolysis and cellulose conversion. The fermentation conditions could be synchronized with hydrolysis optima by carrying out the SSF at a higher temperature, but this would require a thermo-tolerant organism. Economically viable production of platform chemicals from lignocellulosic biomass (LCB) has long been stymied because of the significantly higher cost of hydrolytic enzymes. The major objective of this work is to develop an SSF strategy for D-lactic acid (D-LA) production by a thermo-tolerant organism, in which the enzyme loading could significantly be reduced without compromising on the overall conversion.ResultsA thermo-tolerant strain of Lactobacillus bulgaricus was developed by adaptive laboratory evolution (ALE) which enabled the SSF to be performed at 45 °C with reduced enzyme usage. Despite the reduction of enzyme loading from 15 Filter Paper Unit/gLCB (FPU/gLCB) to 5 FPU/gLCB, we could still achieve?~?8% higher cellulose to D-LA conversion in batch SSF, in comparison to the conversion by separate enzymatic hydrolysis and fermentation processes at 45 °C and pH 5.5. Extending the batch SSF to SSF with pulse-feeding of 5% pre-treated biomass and 5 FPU/gLCB, at 12-h intervals (36th-96th h), resulted in a titer of 108 g/L D-LA and 60% conversion of cellulose to D-LA. This is one among the highest reported D-LA titers achieved from LCB.ConclusionsWe have demonstrated that the SSF strategy, in conjunction with evolutionary engineering, could drastically reduce enzyme requirement and be the way forward for economical production of platform chemicals from lignocellulosics. We have shown that fed-batch SSF processes, designed with multiple pulse-feedings of the pre-treated biomass and enzyme, can be an effective way of enhancing the product concentrations.

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Thermostable Cellulase Biosynthesis from Paenibacillus alvei and its Utilization in Lactic Acid Production by Simultaneous Saccharification and Fermentation.

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