Food and Fermentation Industries >

2023 , Vol. 49 >Issue 9: 1 - 8

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.032922

Endo-β-1,4-glucanase expression in Pichia pastoris for coupled fermentation with Xanthomonas campestris for low-molecular-weight xanthan gum preparation

  • XU Ying ,
  • LIU Zhilei ,
  • ZHAN Xiaobei ,
  • JIANG Yun ,
  • LI Zhitao ,
  • GAO Minjie
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  • (Key Laboratory of Carbohydrate Chemistry and Biotechnology of the Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China)

Received date: 2022-07-09

  Revised date: 2022-07-29

  Online published: 2023-06-05

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Abstract

Low-molecular-weight xanthan gum has various biological activities such as antibacterial, probiotics and antioxidation, so the preparation of low-molecular-weight xanthan gum by hydrolysis method has broad application prospects. The endo-β-1,4-glucanase from sheep rumen metagenome was first expressed in Pichia pastoris, it optimally functioned at pH 7.0 and 75 ℃ in addition to having high pH and temperature stabilities. Thereafter, a coupled fermentation system of P. pastoris and Xanthomonas campestris was established and the fermentation conditions were optimized, endo-β-1,4-glucanase secreted by P. pastoris can directly hydrolyze xanthan gum metabolized by X. campestris to produce functional low-molecular-weight xanthan gum. In a shake flask, the maximum content of total sugar was 11.4 g/L, and the proportion of low-molecular-weight xanthan gum with weight average molecular weight of 2 500 Da reached 93.79%. This study provides a potential strategy for the industrial production of functional low-molecular-weight xanthan gum.

Key words： endo-β-1,4-glucanase; Pichia pastoris; fermentation optimization; coupled fermentation; low-molecular-weight xanthan gum

Cite this article

XU Ying , LIU Zhilei , ZHAN Xiaobei , JIANG Yun , LI Zhitao , GAO Minjie . Endo-β-1,4-glucanase expression in Pichia pastoris for coupled fermentation with Xanthomonas campestris for low-molecular-weight xanthan gum preparation[J]. Food and Fermentation Industries, 2023 , 49(9) : 1 -8 . DOI: 10.13995/j.cnki.11-1802/ts.032922

References

[1] GARCÍA-OCHOA F, SANTOS V E, CASAS J A, et al.Xanthan gum:Production, recovery, and properties[J].Biotechnology Advances, 2000, 18(7):549-579.
[2] JANSSON P E, KENNE L, LINDBERG B.Structure of the extracellular polysaccharide from Xanthomonas campestris[J].Carbohydrate Research, 1975, 45(1):275-282.
[3] OSTROWSKI M P, LA ROSA S L, KUNATH B J, et al.Mechanistic insights into consumption of the food additive xanthan gum by the human gut microbiota[J].Nature Microbiology, 2022, 7(4):556-569.
[4] CHEN Q X, SHAO X T, LING P X, et al.Low molecular weight xanthan gum suppresses oxidative stress-induced apoptosis in rabbit chondrocytes[J].Carbohydrate Polymers, 2017, 169:255-263.
[5] HAN G Y, CHEN Q X, LIU F, et al.Low molecular weight xanthan gum for treating osteoarthritis[J].Carbohydrate Polymers, 2017, 164:386-395.
[6] XU J J, WANG R Y, ZHANG H T, et al.In vitro assessment of prebiotic properties of oligosaccharides derived from four microbial polysaccharides[J].LWT, 2021, 147:111544.
[7] SALEH H M, ANNUAR M S M, SIMARANI K.Ultrasound degradation of xanthan polymer in aqueous solution:Its scission mechanism and the effect of NaCl incorporation[J].Ultrasonics Sonochemistry, 2017, 39:250-261.
[8] LI R S, FEKE D L.Rheological and kinetic study of the ultrasonic degradation of xanthan gum in aqueous solution:Effects of pyruvate group[J].Carbohydrate Polymers, 2015, 124:216-221.
[9] KOOL M M, GRUPPEN H, SWORN G, et al.Comparison of xanthans by the relative abundance of its six constituent repeating units[J].Carbohydrate Polymers, 2013, 98(1):914-921.
[10] LI B, GUO J Q, CHEN W F, et al.Endoxanthanase, a novel β-d-glucanase hydrolyzing backbone linkage of intact xanthan from newly isolated Microbacterium sp.XT11[J].Applied Biochemistry and Biotechnology, 2009, 159(1):24-32.
[11] LIU Z M, NING C, YUAN M X, et al.High-efficiency expression of a superior β-mannanase engineered by cooperative substitution method in Pichia pastoris and its application in preparation of prebiotic mannooligosaccharides[J].Bioresource Technology, 2020, 311:123482.
[12] GAO M J, XU Y, YANG G S, et al.One-step production of functional branched oligoglucosides with coupled fermentation of Pichia pastoris GS115 and Sclerotium rolfsii WSH-G01[J].Bioresource Technology, 2021, 335:125286.
[13] 李雪雁, 武晓尧, 孙春丽, 等.菊芋菊糖粗提液的微生物除杂[J].食品与发酵工业, 2019, 45(5):127-132.
LI X Y, WU X Y, SUN C L, et al.Microbial removal of impurities from Jerusalem artichoke and inulin crude extracts[J].Food and Fermentation Industries, 2019, 45(5):127-132.
[14] 王子朝. 以甘油为底物发酵生产黄原胶及其特性和应用研究[D].无锡:江南大学, 2017.
WANG Z C.Production of xanthan gum with glycerol and the study of its properties and application[D].Wuxi:Jiangnan University, 2017.
[15] ARIAEENEJAD S, SHEYKH ABDOLLAHZADEH MAMAGHANI A, MALEKI M, et al.A novel high performance in-silico screened metagenome-derived alkali-thermostable endo-β-1, 4-glucanase for lignocellulosic biomass hydrolysis in the harsh conditions[J].BMC Biotechnology, 2020, 20(1):56.
[16] LI P Z, ANUMANTHAN A, GAO X G, et al.Expression of recombinant proteins in Pichia pastoris[J].Applied Biochemistry and Biotechnology, 2007, 142(2):105-124.
[17] JAHIC M, WALLBERG F, BOLLOK M, et al.Temperature limited fed-batch technique for control of proteolysis in Pichia pastoris bioreactor cultures[J].Microbial Cell Factories, 2003, 2(1):6.
[18] CELIK E, CALIK P, OLIVER S G.Fed-batch methanol feeding strategy for recombinant protein production by Pichia pastoris in the presence of co-substrate sorbitol[J].Yeast (Chichester, England), 2009, 26(9):473-484.
[19] INAN M, MEAGHER M M.Non-repressing carbon sources for alcohol oxidase (AOX1) promoter of Pichia pastoris[J].Journal of Bioscience and Bioengineering, 2001, 92(6):585-589.
[20] ZHU T C, YOU L J, GONG F Y, et al.Combinatorial strategy of sorbitol feeding and low-temperature induction leads to high-level production of alkaline β-mannanase in Pichia pastoris[J].Enzyme and Microbial Technology, 2011, 49(4):407-412.
[21] 王旺. 毕赤酵母工程菌产植酸酶的发酵优化及放大[D].广州:华南理工大学, 2019.
WANG W.Fermentation optimization and amplification of phytase production by Pichia pastoris engineering bacteria[D].Guangzhou:South China University of Technology, 2019.
[22] 姜海珠, 周海龙, 谷金芸, 等.纤维素酶CtCel8A的异源表达及其降解黄原胶性能[J].大连工业大学学报, 2021, 40(2):79-84.
JIANG H Z, ZHOU H L, GU J Y, et al.Heterologous expression and xanthan-degradation properties of a cellulose CtCel8A[J].Journal of Dalian Polytechnic University, 2021, 40(2):79-84.
[23] 杨国帅, 许颖, 詹晓北, 等.Microbacterium sp.XT11黄原胶内切酶在毕赤酵母中的异源表达、性质及应用[J].食品与发酵工业, 2022, 48(6):8-14.
YANG G S, XU Y, ZHAN X B, et al.Characterization and application of an endoxanthanase from Microbacterium sp.XT11 heterologous expressed in Pichia pastoris[J].Food and Fermentation Industries, 2022, 48(6):8-14.
[24] LIANG Y, ZHU L, DING H, et al.Enhanced production of curdlan by coupled fermentation system of Agrobacterium sp.ATCC 31749 and Trichoderma harzianum GIM 3.442[J].Carbohydrate Polymers, 2017, 157:1 687-1 694.
[25] WU J R, YANG Z L, YANG X C, et al.Synthesis of branched β-1, 3-glucan oligosaccharide with narrow degree of polymerization by fungi co-cultivation[J].Carbohydrate Polymers, 2021, 273:118582.
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