Expression and Characterization of Xyloglucanase from <i>Paenibacillus borealis</i> and Its Application in the Production of Xylogluco-oligosaccharides from Apple Pomace

TIAN Jiaxin; YANG Hang; ZHAO Yuqi; YAN Qiaojuan; LI Yanxiao; JIANG Zhengqiang

doi:10.13386/j.issn1002-0306.2024060437

Turn off MathJax

Article Contents

Abstract

References

TIAN Jiaxin, YANG Hang, ZHAO Yuqi, et al. Expression and Characterization of Xyloglucanase from Paenibacillus borealis and Its Application in the Production of Xylogluco-oligosaccharides from Apple Pomace[J]. Science and Technology of Food Industry, 2025, 46(11): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024060437.

Citation:

PDF (6775 KB)

Expression and Characterization of Xyloglucanase from Paenibacillus borealis and Its Application in the Production of Xylogluco-oligosaccharides from Apple Pomace

1.
Key Laboratory of China National Light Industry and Food Bioengineering, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China
2.
College of Engineering, China Agricultural University, Beijing 100083, China
3.
Food Laboratory of Zhongyuan, Luohe 462300, China
4.
College of Food Science and Engineering, Collaborative Innovation Center for Modern Grain Circulation and Safety, Nanjing University of Finance and Economics, Nanjing 210023, China

More Information

Received Date: June 30, 2024
Available Online: March 30, 2025

Graphical Abstract

Abstract

Abstract

Apple pomace (AP) is rich in xyloglucan, serves as a substrate to produce xylogluco-oligosaccharides with various functional activities. The hydrolysis of AP by novel xyloglucanases is conducive to the high-value utilization of AP. In this study, a novel GH74 xyloglucanase gene (PbXEG74B) from Paenibacillus borealis was cloned and expressed in Escherichia coli BL21 (DE3). The enzyme was characterized and further used to hydrolyze AP for xylogluco-oligosaccharides production. It was expressed in E. coli with an expression level of 18.0 U/mL. The optimal conditions of PbXEG74B were pH5.5 and 55 ℃. It showed good stability in the pH range of 4.5~8.5 and temperatures up to 40 ℃. The enzyme hydrolyzed xyloglucan to produce xylogluco-oligosaccharides with degrees of polymerization (DP) of 2~9. AP was pretreated with deep eutectic solvents (DESs) and hydrolyzed by PbXEG74B to produce xylogluco-oligosaccharides. Under optimal conditions, the yield of xylogluco-oligosaccharides with DP 2~9 was 2.79 g/100 g AP, containing 67.0%, 11.5%, and 21.5% of xylogluco-oligosaccharides with DP 2~4, 5~6, and 7~9, respectively. PbXEG74B with good enzymatic and hydrolytic properties shows great potential in the high value utilization of AP.
- Paenibacillus borealis,
- xyloglucanase,
- apple pomace,
- xylogluco-oligosaccharide,
- deep eutectic solvents

FullText(HTML)

References (41)

References

[1]	ZHU L, QIN S, ZHAI S, et al. Inulin with different degrees of polymerization modulates composition of intestinal microbiota in mice[J]. FEMS Microbiology Letters,2017,364(10):fnx075.
[2]	MUELLER M, REINER J, FLEISCHHACKER L, et al. Growth of selected probiotic strains with fructans from different sources relating to degree of polymerization and structure[J]. Journal of Functional Foods,2016,24:264−275.
[3]	WANG N, LI Y, MIAO M, et al. High level expression of a xyloglucanase from Rhizomucor miehei in Pichia pastoris for production of xyloglucan oligosaccharides and its application in yoghurt[J]. International Journal of Biological Macromolecules,2021,190:845−852.
[4]	CHEN H, JIANG X, LI S, et al. Possible beneficial effects of xyloglucan from its degradation by gut microbiota[J]. Trends in Food Science & Technology,2020,97:65−75.
[5]	LANG W, TAGAMI T, KANG H J, et al. Partial depolymerization of tamarind seed xyloglucan and its functionality toward enhancing the solubility of curcumin[J]. Carbohydrate Polymers,2023,307:120629. doi: 10.1016/j.carbpol.2023.120629
[6]	LI X, CHEN Y, SONG L, et al. Partial enzymolysis affects the digestion of tamarind seed polysaccharides in vitro:Degradation accelerates and gut microbiota regulates[J]. International Journal of Biological Macromolecules,2023,237:124175.
[7]	JULIAN J D, ZABOTINA O A. Xyloglucan biosynthesis:From genes to proteins and their functions[J]. Frontiers in Plant Science,2022,13:920494.
[8]	ATTIA M A, BRUMER H. New family of carbohydrate-binding modules defined by a galactosyl-binding protein module from a Cellvibrio japonicus endo-xyloglucanase[J]. Applied and Environmental Microbiology,2021,87(5):e02634−20.
[9]	SUN P, LI X, DILOKPIMOL A, et al. Fungal glycoside hydrolase family 44 xyloglucanases are restricted to the phylum Basidiomycota and show a distinct xyloglucan cleavage pattern[J]. iScience,2022,25(1):103666.
[10]	VITCOSQUE G L, RIBEIRO L F C, DE LUCAS R C, et al. The functional properties of a xyloglucanase (GH12) of Aspergillus terreus expressed in Aspergillus nidulans may increase performance of biomass degradation[J]. Applied Microbiology and Biotechnology,2016,100:9133−9144.
[11]	BENKŐ Z, SIIKA-AHO M, VIIKARI L, et al. Evaluation of the role of xyloglucanase in the enzymatic hydrolysis of lignocellulosic substrates[J]. Enzyme and Microbial Technology,2008,43(2):109−114. doi: 10.1016/j.enzmictec.2008.03.005
[12]	BAMPIDIS V, AZIMONTI G, MARIA D L B, et al. Safety and efficacy of a feed additive consisting of endo-1,4-β xylanase, endo-1,4-β-glucanase and xyloglucan-specific-endo-β-1,4-glucanase produced by Trichoderma citrinoviride DSM 33578 (Huvezym® neXo) for all Suidae (Huvepharma EOOD)[J]. EFSA Journal,2024,22(3):e8643.
[13]	HSIUNG S Y, LI J, IMRE B, et al. Structures of the xyloglucans in the monocotyledon family Araceae (aroids)[J]. Planta,2023,257(2):39.
[14]	LI K, BARRETT K, AGGER J W, et al. Bioinformatics-based identification of GH12 endoxyloglucanases in citrus-pathogenic Penicillium spp[J]. Enzyme and Microbial Technology,2024,178:110441.
[15]	DAMÁSIO A R L, RIBEIRO L F C, RIBEIRO L F, et al. Functional characterization and oligomerization of a recombinant xyloglucan-specific endo-β-1,4-glucanase (GH12) from Aspergillus niveus[J]. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics,2012,1824(3):461−467. doi: 10.1016/j.bbapap.2011.12.005
[16]	ARNAL G, STOGIOS P J, ASOHAN J, et al. Substrate specificity, regiospecificity, and processivity in glycoside hydrolase family 74[J]. Journal of Biological Chemistry,2019,294(36):13233−13247.
[17]	GUARDIA L, SUÁREZ L, QUEREJETA N, et al. Apple waste:A sustainable source of carbon materials and valuable compounds[J]. ACS Sustainable Chemistry & Engineering,2019,7(20):17335−17343.
[18]	CHEN M, MAC-BÉAR J, ROPARTZ D, et al. Biorefinery of apple pomace:New insights into xyloglucan building blocks[J]. Carbohydrate Polymers,2022,290:119526.
[19]	FULLERTON C G, PRAKASH R, NINAN A S, et al. Fruit from two kiwifruit genotypes with contrasting softening rates show differences in the xyloglucan and pectin domains of the cell wall[J]. Frontiers in Plant Science,2020,11:537386.
[20]	田锐, 胡亚洁, 刘巧玲, 等. 低共熔溶剂在木质纤维生物质预处理领域的研究进展[J]. 中国造纸,2022,41(3):78−86. [TIAN Rui, HU Yajie, LIU Qiaoling, et al. Research progress of deep eutectic solvents in Lignocellulosic biomass pretreatment[J]. China Pulp & Paper,2022,41(3):78−86.] doi: 10.11980/j.issn.0254-508X.2022.03.011 TIAN Rui, HU Yajie, LIU Qiaoling, et al. Research progress of deep eutectic solvents in Lignocellulosic biomass pretreatment[J]. China Pulp & Paper, 2022, 41（3）: 78−86. doi: 10.11980/j.issn.0254-508X.2022.03.011
[21]	CHEN M, FALOURD X, LAHAYE M. Sequential natural deep eutectic solvent pretreatments of apple pomace:A novel way to promote water extraction of pectin and to tailor its main structural domains[J]. Carbohydrate Polymers,2021,266:118113.
[22]	ELO S, SUOMINEN I, KÄMPFER P, et al. Paenibacillus borealis sp. nov., a nitrogen-fixing species isolated from spruce forest humus in Finland[J]. International Journal of Systematic and Evolutionary Microbiology,2001,51(2):535−545.
[23]	JUMPER J, EVANS R, PRITZEL A, et al. Highly accurate protein structure prediction with AlphaFold[J]. Nature,2021,596(7873):583−589. doi: 10.1038/s41586-021-03819-2
[24]	MILLER C L. Use of dinitrosalycylic acid reagent for determination of reducing sugar[J]. Analytical Chemistry,1959,31(3):426−428.
[25]	CLASSICS LOWRY O, ROSEBROUGH N, FARR A, et al. Protein measurement with the Folin phenol reagent[J]. J Biol Chem,1951,193(1):265−75.
[26]	SATO S, OHTA K, KOJIMA K, et al. Isolation and characterization of two types of xyloglucanases from a phytopathogenic fungus, Verticillium dahliae[J]. Journal of Applied Glycoscience,2016,63(1):13−18.
[27]	GRISHUTIN S G, GUSAKOV A V, MARKOV A V, et al. Specific xyloglucanases as a new class of polysaccharide-degrading enzymes[J]. Biochimica et Biophysica Acta (BBA)-General Subjects,2004,1674(3):268−281.
[28]	RYKOV S V, KORNBERGER P, HERLET J, et al. Novel endo-(1,4)-β-glucanase Bgh12A and xyloglucanase Xgh12B from Aspergillus cervinus belong to GH12 subgroup I and II, respectively[J]. Applied Microbiology and Biotechnology,2019,103:7553−7566.
[29]	POL D, MENON V, RAO M. Biochemical characterization of a novel thermostable xyloglucanase from an alkalothermophilic Thermomonospora sp[J]. Extremophiles,2012,16:135−146.
[30]	SIANIDIS G, POZIDIS C, BECKER F, et al. Functional large-scale production of a novel Jonesia sp. xyloglucanase by heterologous secretion from Streptomyces lividans[J]. Journal of Biotechnology,2006,121(4):498−507.
[31]	YAOI K, NAKAI T, KAMEDA Y, et al. Cloning and characterization of two xyloglucanases from Paenibacillus sp. strain KM21[J]. Applied and Environmental Microbiology,2005,71(12):7670−7678.
[32]	SHI H, GUO J, YAN X, et al. Characterization of a xyloglucananse in biodegradation of woody plant xyloglucan from Caldicellulosiruptor kronotskyensis[J]. BioResources,2022,17(1):673.
[33]	WANG N, LI Y, ZHU C, et al. High-level expression of xyloglucanase B from Rhizomucor miehei and its application in the preparation of partially hydrolyzed apple pomace xyloglucan[J]. Food Bioengineering,2022,1(2):119−125.
[34]	ICHINOSE H, ARAKI Y, MICHIKAWA M, et al. Characterization of an endo-processive-type xyloglucanase having a β-1, 4-glucan-binding module and an endo-type xyloglucanase from Streptomyces avermitilis[J]. Applied and Environmental Microbiology,2012,78(22):7939−7945.
[35]	MATSUZAWA T, KAMEYAMA A, NAKAMICHI Y, et al. Identification and characterization of two xyloglucan-specific endo-1,4-glucanases in Aspergillus oryzae[J]. Applied Microbiology and Biotechnology,2020,104:8761−8773.
[36]	ARNAL G, STOGIOS P J, ASOHAN J, et al. Structural enzymology reveals the molecular basis of substrate regiospecificity and processivity of an exemplar bacterial glycoside hydrolase family 74 endo-xyloglucanase[J]. Biochemical Journal,2018,475(24):3963−3978.
[37]	BEREZINA O V, HERLET J, RYKOV S V, et al. Thermostable multifunctional GH74 xyloglucanase from Myceliophthora thermophila:High-level expression in Pichia pastoris and characterization of the recombinant protein[J]. Applied Microbiology and Biotechnology,2017,101:5653−5666.
[38]	DAMASIO A R L, RUBIO M V, GONÇALVES T A, et al. Xyloglucan breakdown by endo-xyloglucanase family 74 from Aspergillus fumigatus[J]. Applied Microbiology and Biotechnology,2017,101:2893−2903. doi: 10.1007/s00253-016-8014-6
[39]	POWLOWSKI J, MAHAJAN S, SCHAPIRA M, et al. Substrate recognition and hydrolysis by a fungal xyloglucan-specific family 12 hydrolase[J]. Carbohydrate Research,2009,344(10):1175−1179.
[40]	WANG B, CHEN K, ZHANG P, et al. Comparison of the biochemical properties and roles in the xyloglucan-rich biomass degradation of a GH74 xyloglucanase and its CBM-deleted variant from Thielavia terrestris[J]. International Journal of Molecular Sciences,2022,23(9):5276.
[41]	SHI R, YANG S, WANG N, et al. Synthesis of 2′-fucosyllactose from apple pomace-derived xyloglucan oligosaccharides by an α-L-fucosidase from Pedobacter sp. CAU209[J]. Applied Microbiology and Biotechnology,2023,107(11):3579−3591.