Identification and Immobilization of an Invertase With High Specific Activity and Sucrose Tolerance Ability of Sp. W5 for High Fructose Syrup Preparation

Overview

Journal Front Microbiol

Specialty Microbiology

Date 2020 Apr 25

PMID 32328053

Citations 8

Authors

Gang Zhou

Can Peng

Xiaosa Liu

Fei Chang

Yazhong Xiao

Juanjuan Liu

Zemin Fang

Affiliations

Soon will be listed here.

Abstract

Invertases catalyze the hydrolysis of sucrose into fructose and glucose and can be employed as an alternative in producing high fructose syrup. In this study, we reported the heterologous expression of an invertase gene () of sp. w5 in . GspInv activity reached 147.6 ± 0.4 U/mL after 5 days of methanol induction. GspInv is invertase with a high specific activity of 2,776.1 ± 124.2 U/mg toward sucrose. GspInv showed high tolerance to sucrose ( = 1.2 M), glucose ( > 2 M), fructose ( = 1.5 M), and a variety of metal ions that make it an ideal candidate for high fructose syrup production. A carbohydrate-binding module was sequence-optimized and fused to the N-terminus of GspInv. The fusion protein had the highest immobilization efficiency at room temperature within 1 h adsorption, with 1 g of cellulose absorption up to 8,000 U protein. The cellulose-immobilized fusion protein retained the unique properties of GspInv. When applied in high fructose syrup preparation by using 1 M sucrose as the substrate, the sucrose conversion efficiency of the fused protein remained at approximately 95% after 50 h of continuous hydrolysis on a packed bed reactor. The fused protein can also hydrolyze completely the sucrose in sugarcane molasses. Our results suggest that GspInv is an unusual invertase and a promising candidate for high fructose syrup preparation.

Citing Articles

Unveiling species diversity within early-diverging fungi from China III: Six new species and a new record of (Cunninghamellaceae, Mucoromycota).

Wang Y, Zhao H, Jiang Y, Liu X, Tao M, Liu X MycoKeys. 2024; 110:287-317.

PMID: 39610859 PMC: 11603104. DOI: 10.3897/mycokeys.110.130260.

Industrial Approach to Invertase Production from Fruit Waste for Enhanced Efficiency and Conservation.

Dokuzparmak E ACS Omega. 2024; 9(24):26183-26194.

PMID: 38911758 PMC: 11190939. DOI: 10.1021/acsomega.4c01732.

Expression and characterization of a protease-resistant β-d-fructofuranosidase BbFFase9 gene suitable for preparing invert sugars from soybean meal.

Chen Z, Shen Y, Wang R, Li S, Jia Y Heliyon. 2023; 9(9):e19889.

PMID: 37809427 PMC: 10559283. DOI: 10.1016/j.heliyon.2023.e19889.

A Strategy for Rapid Acquisition of the β-D-Fructofuranosidase Gene through Chemical Synthesis and New Function of Its Encoded Enzyme to Improve Gel Properties during Yogurt Processing.

Chen Z, Shen Y, Xu J Foods. 2023; 12(8).

PMID: 37107499 PMC: 10137638. DOI: 10.3390/foods12081704.

Efficient Degradation for Raffinose and Stachyose of a β-D-Fructofuranosidase and Its New Function to Improve Gel Properties of Coagulated Fermented-Soymilk.

Chen Z, Shen Y, Xu J Gels. 2023; 9(4).

PMID: 37102957 PMC: 10137817. DOI: 10.3390/gels9040345.

References

Hussack G, Luo Y, Veldhuis L, Hall J, Tanha J, MacKenzie R . Multivalent anchoring and oriented display of single-domain antibodies on cellulose. Sensors (Basel). 2012; 9(7):5351-67. PMC: 3274147. DOI: 10.3390/s90705351. View

Shoseyov O, Shani Z, Levy I . Carbohydrate binding modules: biochemical properties and novel applications. Microbiol Mol Biol Rev. 2006; 70(2):283-95. PMC: 1489539. DOI: 10.1128/MMBR.00028-05. View

Tabuchi S, Ito J, Adachi T, Ishida H, Hata Y, Okazaki F . Display of both N- and C-terminal target fusion proteins on the Aspergillus oryzae cell surface using a chitin-binding module. Appl Microbiol Biotechnol. 2010; 87(5):1783-9. PMC: 2903697. DOI: 10.1007/s00253-010-2664-6. View

Hsieh C, Liu L, Yeh S, Chen C, Lin H, Sung H . Molecular cloning and functional identification of invertase isozymes from green bamboo Bambusa oldhamii. J Agric Food Chem. 2006; 54(8):3101-7. DOI: 10.1021/jf052711s. View

Acosta N, Beldarrain A, Rodriguez L, Alonso Y . Characterization of recombinant invertase expressed in methylotrophic yeasts. Biotechnol Appl Biochem. 2000; 32(3):179-87. DOI: 10.1042/ba20000064. View

Zhou J, He L, Gao Y, Han N, Zhang R, Wu Q . Characterization of a novel low-temperature-active, alkaline and sucrose-tolerant invertase. Sci Rep. 2016; 6:32081. PMC: 4995436. DOI: 10.1038/srep32081. View

Singh R, Chauhan K, Pandey A, Larroche C, Kennedy J . Purification and characterization of two isoforms of exoinulinase from Penicillium oxalicum BGPUP-4 for the preparation of high fructose syrup from inulin. Int J Biol Macromol. 2018; 118(Pt B):1974-1983. DOI: 10.1016/j.ijbiomac.2018.07.040. View

Goradia D, Cooney J, Hodnett B, Magner E . Characteristics of a mesoporous silicate immobilized trypsin bioreactor in organic media. Biotechnol Prog. 2006; 22(4):1125-31. DOI: 10.1021/bp050334y. View

Liu Y, Zheng P, Sun Z, Ni Y, Dong J, Zhu L . Economical succinic acid production from cane molasses by Actinobacillus succinogenes. Bioresour Technol. 2007; 99(6):1736-42. DOI: 10.1016/j.biortech.2007.03.044. View

10.

Singh R, Chauhan K, Pandey A, Larroche C . Biocatalytic strategies for the production of high fructose syrup from inulin. Bioresour Technol. 2018; 260:395-403. DOI: 10.1016/j.biortech.2018.03.127. View

11.

Wang L, Wang A, Hsieh C, Chen C, Sung H . Vacuolar invertases in sweet potato: molecular cloning, characterization, and analysis of gene expression. J Agric Food Chem. 2005; 53(9):3672-8. DOI: 10.1021/jf0480851. View

12.

Cai S, Xiong Z, Li L, Li M, Zhang L, Liu C . Differential responses of root growth, acid invertase activity and transcript level to copper stress in two contrasting populations of Elsholtzia haichowensis. Ecotoxicology. 2013; 23(1):76-91. DOI: 10.1007/s10646-013-1153-y. View

13.

Lu L, Xu S, Zhao R, Zhang D, Li Z, Li Y . Synthesis of galactooligosaccharides by CBD fusion β-galactosidase immobilized on cellulose. Bioresour Technol. 2012; 116:327-33. DOI: 10.1016/j.biortech.2012.03.108. View

14.

Veana F, Fuentes-Garibay J, Aguilar C, Rodriguez-Herrera R, Guerrero-Olazaran M, Viader-Salvado J . Gene encoding a novel invertase from a xerophilic Aspergillus niger strain and production of the enzyme in Pichia pastoris. Enzyme Microb Technol. 2014; 63:28-33. DOI: 10.1016/j.enzmictec.2014.05.001. View

15.

Xia J, Xu J, Hu L, Liu X . Enhanced poly(L-malic acid) production from pretreated cane molasses by Aureobasidium pullulans in fed-batch fermentation. Prep Biochem Biotechnol. 2016; 46(8):798-802. DOI: 10.1080/10826068.2015.1135464. View

16.

Finn R, Attwood T, Babbitt P, Bateman A, Bork P, Bridge A . InterPro in 2017-beyond protein family and domain annotations. Nucleic Acids Res. 2016; 45(D1):D190-D199. PMC: 5210578. DOI: 10.1093/nar/gkw1107. View

17.

Nadeem H, Rashid M, Riaz M, Asma B, Javed M, Perveen R . Invertase from hyper producer strain of Aspergillus niger: physiochemical properties, thermodynamics and active site residues heat of ionization. Protein Pept Lett. 2009; 16(9):1098-105. DOI: 10.2174/092986609789055322. View

18.

Trollope K, Gorgens J, Volschenk H . Semirational Directed Evolution of Loop Regions in Aspergillus japonicus β-Fructofuranosidase for Improved Fructooligosaccharide Production. Appl Environ Microbiol. 2015; 81(20):7319-29. PMC: 4579456. DOI: 10.1128/AEM.02134-15. View

19.

Meng H, Zhang Z, Chen M, Su Y, Li L, Miyoshi S . Characterization and horizontal transfer of class 1 integrons in Salmonella strains isolated from food products of animal origin. Int J Food Microbiol. 2011; 149(3):274-7. DOI: 10.1016/j.ijfoodmicro.2011.07.006. View

20.

van Wyk N, Trollope K, Steenkamp E, Wingfield B, Volschenk H . Identification of the gene for β-fructofuranosidase from Ceratocystis moniliformis CMW 10134 and characterization of the enzyme expressed in Saccharomyces cerevisiae. BMC Biotechnol. 2013; 13:100. PMC: 3880211. DOI: 10.1186/1472-6750-13-100. View