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Using One-pot Fermentation Technology to Prepare Enzyme Cocktail to Sustainably Produce Low Molecular Weight Galactomannans from Sesbania cannabina Seeds

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Abstract

Enzymatic hydrolysis using β-mannanase and α-galactosidase is necessary to produce low molecular weight galactomannan (LMW-GM) from galactomannans (GM) in the leguminous seeds. In this study, different ratios of avicel and melibiose were used as the inductors (carbon sources) for Trichoderma reesei to metabolize the enzyme cocktail containing β-mannanase and α-galactosidase using one-pot fermentation technology. The obtained enzyme cocktail was used to efficiently produce LMW-GM from GM in Sesbania cannabina seeds. Results showed that 15 g/L avicel and 10 g/L melibiose were the best carbon sources to prepare enzyme cocktail containing β-mannanase and α-galactosidase with activities of 3.69 ± 0.27 U/mL and 0.51 ± 0.02 U/mL, respectively. Specifically, melibiose could effectively induce the metabolite product of α-galactosidase by T. reesei, which showed good performance in degrading the galactose substituent from GM backbone. The degradation of galactose alleviated the spatial site-blocking effect for enzymatic hydrolysis by β-mannanase and improved the yield of LMW-GM. This research can lay the foundation for the industrial technology amplification of LMW-GM production for further application.

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Data Availability

Data and material are available upon reasonable request to the corresponding author.

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Abbreviations

LMW-GM:

low molecular weight galactomannans

GM:

galactomannans

T. reesei :

Trichoderma reesei

LMWP:

low molecular weight polysaccharides

pNPM:

β-D-Mannopyranoside,4-nitrophenyl

pNPG:

p-nitrophenyl-β-D-galactopyranoside

LBG:

locust bean gum

DNS:

3,5-dinitrosalicylic acid

PNP:

p-nitrophenol spectral profile

GMOS:

galactomannan oligosaccharides

HPSEC:

high-performance size exclusion chromatography

Mw:

weight average molecular weight

Mn:

number average molecular weight

PDI:

polydispersity index

DPw:

weight average degree of polymerization

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Funding

This work was supported by the National Key R&D Program of China (2016YFD0600803) and Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Authors and Affiliations

  1. Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China

    Bowen Yan, Yuheng Tao, Caoxing Huang, Chenhuan Lai & Qiang Yong

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  1. Bowen Yan
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  2. Yuheng Tao
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  3. Caoxing Huang
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  4. Chenhuan Lai
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CXH proposed the idea. BWY and YHT performed the experiments and wrote the manuscript. CHL and QY wrote and revised the manuscript. All the authors contributed to write the manuscript. All authors read and approved the final manuscript.

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Correspondence to Qiang Yong.

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Yan, B., Tao, Y., Huang, C. et al. Using One-pot Fermentation Technology to Prepare Enzyme Cocktail to Sustainably Produce Low Molecular Weight Galactomannans from Sesbania cannabina Seeds. Appl Biochem Biotechnol 194, 3016–3030 (2022). https://doi.org/10.1007/s12010-022-03891-y

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