Improving the thermostability and catalytic efficiency of GH11 xylanase PjxA by adding disulfide bridges

Chao Teng; Yuefeng Jiang; Youqiang Xu; Qin Li; Xiuting Li; Guangsen Fan; Ke Xiong; Ran Yang; Chengnan Zhang; Rong Ma; Yunping Zhu; Jinlong Li; Changtao Wang

doi:10.1016/j.ijbiomac.2019.01.087

Improving the thermostability and catalytic efficiency of GH11 xylanase PjxA by adding disulfide bridges

Int J Biol Macromol. 2019 May 1:128:354-362. doi: 10.1016/j.ijbiomac.2019.01.087. Epub 2019 Jan 22.

Authors

Chao Teng¹, Yuefeng Jiang², Youqiang Xu³, Qin Li⁴, Xiuting Li⁵, Guangsen Fan³, Ke Xiong², Ran Yang², Chengnan Zhang³, Rong Ma⁶, Yunping Zhu³, Jinlong Li³, Changtao Wang⁷

Affiliations

¹ Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing 100048, China; Beijing Engineering and Technology Research Center of Food Additives, Beijing 100048, China; School of Food and Chemical Engineering, Beijing Technology and Business University, No. 11, Fucheng Road, Beijing 100048, China.
² Beijing Engineering and Technology Research Center of Food Additives, Beijing 100048, China; School of Food and Chemical Engineering, Beijing Technology and Business University, No. 11, Fucheng Road, Beijing 100048, China.
³ School of Food and Chemical Engineering, Beijing Technology and Business University, No. 11, Fucheng Road, Beijing 100048, China.
⁴ School of Food and Biological Engineering, Hubei University of Technology, No. 28, Nanli Road, Wuhan 430068, China.
⁵ Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing 100048, China; Beijing Engineering and Technology Research Center of Food Additives, Beijing 100048, China; School of Food and Chemical Engineering, Beijing Technology and Business University, No. 11, Fucheng Road, Beijing 100048, China. Electronic address: lixt@btbu.edu.cn.
⁶ School of Foreign Language, Beijing Technology and Business University, No. 33, Fucheng Road, Beijing 100048, China.
⁷ Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing 100048, China; School of Science, Beijing Technology and Business University, No. 11, Fucheng Road, Beijing 100048, China. Electronic address: Wangct@btbu.edu.cn.

PMID: 30682487
DOI: 10.1016/j.ijbiomac.2019.01.087

Abstract

In order to increase the thermostability and catalytic efficiency of acidophilic GH11 xylanase, two disulfide bonds were introduced into crucial region of the enzyme. The xylanase PjxA, from Penicillium janthinellum MA21601, has attracted considerable attention due to its favorable acid-resistance; however, its poor thermostability and low enzymatic hydrolysis efficiency limit its application. In this study, two disulfide bonds were introduced into crucial regions of three recombined xylanases (DB-s1s3, DB-s1s4, and DB-s3s4). All three xylanases remained acid-resistant while gaining improved hydrolytic and thermostability properties, of which DB-s1s3 was the most noteworthy. The optimal temperature of recombined xylanase DB-s1s3 increased from 50 °C to 70 °C. The specific activity of DB-s1s3 was 4.76-fold higher than that of wild-type xylanase PjxA. Moreover, DB-s1s3 showed a 2.14-fold increase in k_cat/K_m. In addition, DB-s1 s3 showed improved hydrolytic characteristics, of which the most noteworthy was its enhanced ability to produce xylose and xylobiose from polymeric substrates. Compared with PjxA, combining DB-s1 s3 with cellulase improved the hydrolytic yield of corncob powder, procuring concentrations of xylose (X1) and xylobiose (X2) of 204.4% and 24.4%, respectively. Thus, these mutants offer great potential for application in the agricultural residue degradation industry.

Keywords: Catalytic characteristics; Disulfide bridge; Xylanase.

MeSH terms

Biocatalysis*
Disulfides / chemistry*
Endo-1,4-beta Xylanases / chemistry*
Endo-1,4-beta Xylanases / genetics
Endo-1,4-beta Xylanases / metabolism*
Enzyme Stability / genetics
Hydrogen-Ion Concentration
Hydrolysis
Kinetics
Models, Molecular
Mutation
Penicillium / enzymology
Protein Conformation
Protein Engineering*
Substrate Specificity
Temperature*
Zea mays / chemistry

Substances

Disulfides
Endo-1,4-beta Xylanases