Optimization of the Fermentation Conditions of <i>Bacillus subtilis</i> Z-5 Producing Pectinase and Its Application

ZHANG Xiaodan; GAO Mengdi; ZHAO Saisai; LIU Keyu; NING Xibin

doi:10.13386/j.issn1002-0306.2022120061

Volume 44 Issue 19

Oct. 2023

Turn off MathJax

Article Contents

Abstract

References

Science and Technology of Food Industry > 2023 > 44(19): 117-127. > DOI: 10.13386/j.issn1002-0306.2022120061

ZHANG Xiaodan, GAO Mengdi, ZHAO Saisai, et al. Optimization of the Fermentation Conditions of Bacillus subtilis Z-5 Producing Pectinase and Its Application[J]. Science and Technology of Food Industry, 2023, 44(19): 117−127. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022120061.

Citation:

PDF (2173 KB)

Optimization of the Fermentation Conditions of Bacillus subtilis Z-5 Producing Pectinase and Its Application

1.
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
2.
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai 201306, China
3.
National R&D Branch Center for Freshwater Aquatic Products Processing Technology (Shanghai), Shanghai 201306, China

More Information

Received Date: December 07, 2022
Available Online: August 04, 2023

Graphical Abstract

Abstract

Abstract

To further improve the yield of pectinase, the fermentation medium and fermentation conditions for pectinase production by Bacillus subtilis Z-5 were optimized using the response surface methodology, and the optimized pectinase was applied for the clarification of pear juice. Based on the single-factor experiments, the Plackett-Burman test design was applied to screen out three significant factors affecting enzyme production: Yeast powder content, pH and incubation time, and then combined with the response surface design method to determine the optimal fermentation conditions. The results showed that the optimal fermentation medium composition was 10 g/L pectin, 7.99 g/L yeast powder and 0.5 g/L MgSO₄·7H₂O, the optimal fermentation conditions were initial pH5.66, fermentation temperature 37 ℃, incubation time 72 h, inoculum 6% (v/v), liquid volume 50/250 mL and substrate concentration 10 g/L. On the basis of this, the enzymatic activity of pectinase produced by B. subtilis Z-5 was increased from 879.00 U/mL to 1549.62 U/mL, which was 1.76 times higher than the unoptimised one. In pear juice clarification experiments, it was shown that the optimum addition of pectinase was 4 mL, the optimum enzymatic time was 2.5 h, the optimum enzymatic temperature was 65 ℃ and the optimum enzymatic pH was 6.0, the maximum light transmission rate was 79.77%. Compared with the commercial enzyme, the homemade pectinase is a compound enzyme with multiple enzyme systems, which can act on other components such as protein and starch contained in the pear juice, making the pear juice more clarified. The results of this study have provided theoretical support for the industrial production and application of pectinase.
- pectinase,
- Bacillus subtilis Z-5,
- optimization of fermentation conditions,
- fruit juice clarification

FullText(HTML)

References (24)

References

[1]	TAKCI H A M, TURKMEN F U. Extracellular pectinase production and purification from a newly isolated Bacillus subtilis strain[J]. International Journal of Food Properties,2016,19(11):2443−2450. doi: 10.1080/10942912.2015.1123270
[2]	JOSHI M, NERURKAR M, ADIVAREKAR R. Characterization, kinetic, and thermodynamic studies of marine pectinase from Bacillus subtilis[J]. Preparative Biochemistry and Biotechnology,2015,45(3):205−220. doi: 10.1080/10826068.2014.907181
[3]	KAUR J S, GUPTA K V. Production of pectinolytic enzymes pectinase and pectin lyase by Bacillus subtilis SAV-21 in solid state fermentation[J]. Annals of Microbiology,2017,67(4):333−342. doi: 10.1007/s13213-017-1264-4
[4]	张翠. 根霉液体发酵生产碱性果胶酶及应用研究[D]. 无锡: 江南大学, 2009. ZHANG C. Study on production of basic pectinase by liquid fermentation of Rhizopus sp.[D]. Wuxi: Jiangnan University, 2009.
[5]	程红. 黑曲霉高产果胶酶菌株YY-21选育、固态发酵工艺及其在沙果汁加工中的应用[D]. 齐齐哈尔: 齐齐哈尔大学, 2013. CHENG H. Selection and breeding of Aspergillus niger high-yielding pectinase strain YY-21, solid-state fermentation process and its application in sand juice processing[D]. Qiqihaer: Qiqihar University, 2013.
[6]	吕丽娟. 扩展青霉YL-9利用苹果渣发酵产果胶酶的研究[D]. 咸阳: 西北农林科技大学, 2011. LÜ L J. Study on the production of pectinase by fermentation of apple pomace using extended Penicillium yl-9[D]. Xianyang: Northwest A and F University, 2011.
[7]	汤鸣强. 黑曲霉产果胶酶的分离纯化和酶学特性研究[D]. 福州: 福建师范大学, 2004. TANG M Q. Isolation, purification and enzymatic characterization of pectinase from Aspergillus niger[D]. Fuzhou: Fujian Normal University, 2014.
[8]	张浩森. 果胶酶高产菌种的筛选及其酶学性质的研究[D]. 无锡: 江南大学, 2008. ZHANG H S. Screening of high-yielding pectinase strains and study of their enzymatic properties[D]. Wuxi: Jiangnan University, 2008.
[9]	杨同香, 吴孔阳, 白云飞, 等. 微生物果胶酶的研究进展[J]. 食品与机械,2020,36(8):201−209. [YANG T X, WU K Y, BAI Y F, et al. Advances in microbial pectinase research[J]. Food and Machinery,2020,36(8):201−209. YANG T X, WU K Y, BAI Y F, et al. Advances in microbial pectinase research[J]. Food and Machinery, 2020, 36(8): 201 - 209.
[10]	张铭, 宁少华, 高茜, 等. 响应面法优化森吉木霉M75菌株发酵培养条件[J]. 林业科学研究,2022,35(5):196−205. [ZHANG M, NING S H, GAO Q, et al. Optimization of fermentation conditions of Trichoderma songyi M75 strain by response surface methodology[J]. Forest Research,2022,35(5):196−205. ZHANG M, NING S H, GAO Q, et al. Optimization of fermentation conditions of Trichoderma songyi M75 strain by response surface methodology[J]. Forest Research, 2022, 35(5): 196 - 205.
[11]	白兰芳, 高慧, 刘明启, 等. 产果胶酶枯草芽孢杆菌的鉴定发酵条件优化及产物酶学性质的研究[J]. 中国畜牧杂志,2011,47(19):63−68. [BAI L F, GAO H, LIU M Q, et al. Optimization of fermentation conditions for the identification of pectinase producing Bacillus subtilis and study of the enzymatic properties of the product[J]. Chinese Journal of Animal Science,2011,47(19):63−68. BAI L F, GAO H, LIU M Q, et al. Optimization of fermentation conditions for the identification of pectinase producing Bacillus subtilis and study of the enzymatic properties of the product[J]. Chinese Journal of Animal Science, 2011, 47(19): 63 - 68.
[12]	GUPTA S, KAPOOR M, SHARMA K K, et al. Production and recovery of an alkaline exo-polygalacturonase from Bacillus subtilis RCK under solid-state fer-mentation using statistical approach[J]. Bioresour Technol,2008,99(5):937−945. doi: 10.1016/j.biortech.2007.03.009
[13]	廖敏, 陈希文, 姜立春, 等. 假单胞菌B41产果胶酶发酵条件的研究[J]. 中国酿造,2012,31(11):58−62. [LIAO M, CHEN X W, JIANG L C, et al. Study of fermentation conditions for pectinase production by Pseudomonas aeruginosa B41[J]. China Brewing,2012,31(11):58−62. doi: 10.3969/j.issn.0254-5071.2012.11.015 LIAO M, CHEN X W, JIANG L C, et al. Study of fermentation conditions for pectinase production by Pseudomonas aeruginosa B41[J]. China Brewing, 2012, 31(11): 58 - 62. doi: 10.3969/j.issn.0254-5071.2012.11.015
[14]	刘均珠, 黄玉婷, 梁丽陈, 等. 产果胶酶菌株的筛选鉴定及产酶条件初步优化[J]. 饲料研究,2022,45(11):71−75. [LIU J Z, HUANG Y T, LIANG L C, et al. Screening and identification of pectinase producing strains and preliminary optimization of enzyme production conditions[J]. Feed Research,2022,45(11):71−75. LIU J Z, HUANG Y T, LIANG L C, et al. Screening and identification of pectinase producing strains and preliminary optimization of enzyme production conditions[J]. Feed Research, 2022, 45(11): 71 - 75.
[15]	吕秀红. 产耐热型酸性果胶酶菌株的筛选、发酵条件优化及酶学性质的研究[D]. 杭州: 浙江工商大学, 2016. LÜ X H. Screening of strains producing heat-resistant acid pectinase, optimization of fermentation conditions and study of enzymatic properties[D]. Hangzhou: Zhejiang Gongshang University, 2016.
[16]	蓝丽精. 高产果胶酶菌株的筛选、发酵条件优化及其应用的研究[D]. 杭州: 浙江师范大学, 2012. LAN L J. Screening of pectinase producing strains, optimization of fermentation conditions and their application[D]. Hangzhou: Zhejiang Normal University, 2012.
[17]	王小敏, 吴文龙, 闾连飞, 等. 分光光度计法测定果胶酶活力的方法研究[J]. 食品工业科技,2007(5):227−229. [WANG X M, WU W L, LÜ L F, et al. Study on determination of pectinase activity by spectrophotometer[J]. Science and Technology of Food Industry,2007(5):227−229. WANG X M, WU W L, LÜ L F, et al. Study on determination of pectinase activity by spectrophotometer[J]. Science and Technology of Food Industry, 2007(5): 227 - 229.
[18]	张飞, 岳田利, 费坚, 等. 果胶酶活力的测定方法研究[J]. 西北农业学报,2004(4):134−137. [ZHANG F, YUE T L, FEI J, et al. Study on determination method of pectinase activity[J]. Acta Agriculturae Boreali-occidentalis Sinica,2004(4):134−137. doi: 10.3969/j.issn.1004-1389.2004.04.030 ZHANG F, YUE T L, FEI J, et al. Study on determination method of pectinase activity[J]. Acta Agriculturae Boreali-occidentalis Sinica, 2004(4): 134 - 137. doi: 10.3969/j.issn.1004-1389.2004.04.030
[19]	谷艺明. 果胶酶高产菌株的筛选、发酵优化及酶学性质的研究[D]. 常州: 常州大学, 2021. GU Y M. Screening, fermentation optimization and enzymatic properties of pectinase producing strains[D]. Changzhou: Changzhou University, 2021.
[20]	顾燕松. 纺织生物助剂果胶酶酶活的测定方法[J]. 纺织科学研究,2002(3):29−35. [GU Y S. Method for determination of enzymatic activity of textile biological auxiliaries pectinase[J]. Textile Science Research,2002(3):29−35. GU Y S. Method for determination of enzymatic activity of textile biological auxiliaries pectinase[J]. Textile Science Research, 2002(3): 29 - 35.
[21]	于平, 王欣馨, 任倩, 等. 产碱性果胶酶菌株的筛选和鉴定及其酶学性质[J]. 中国食品学报,2018,18(9):288−296. [YU P, WANG X X, REN Q, et al. Screening and identification of alkaline pectinase producing strains and their enzymatic properties[J]. Journal of Chinese Institute of Food Science and Technology,2018,18(9):288−296. doi: 10.16429/j.1009-7848.2018.09.037 YU P, WANG X X, REN Q, et al. Screening and identification of alkaline pectinase producing strains and their enzymatic properties[J]. Journal of Chinese Institute of Food Science and Technology, 2018, 18(9): 288 - 296. doi: 10.16429/j.1009-7848.2018.09.037
[22]	陈成, 宁喜斌. 响应面法优化产β-内酰胺酶菌Bacillus cereus B03的发酵条件[J]. 食品工业科技,2020,41(10):138−145. [CHEN C, NING X B. Optimization of fermentation conditions of β-lactamase producing bacteria Bacillus cereus B03 by response surface methodology[J]. Science and Technology of Food Industry,2020,41(10):138−145. CHEN C, NING X B. Optimization of fermentation conditions of β-lactamase producing bacteria Bacillus cereus B03 by response surface methodology[J]. Science and Technology of Food Industry, 2020, 41(10): 138 - 145.
[23]	于平, 吴赟婷, 杨柳贞, 等. 蜡样芽孢杆菌高产新型中性蛋白酶发酵条件的优化[J]. 中国食品学报,2020,20(1):109−117. [YU P, WU Y T, YANG L Z, et al. Optimization of fermentation conditions for high yield of a new neutral protease from Bacillus cereus[J]. Journal of Chinese Institute of Food Science and Technology,2020,20(1):109−117. YU P, WU Y T, YANG L Z, et al. Optimization of fermentation conditions for high yield of a new neutral protease from Bacillus cereus[J]. Journal of Chinese Institute of Food Science and Technology, 2020, 20(1): 109 – 117.
[24]	阿衣满古力·哈力哈拜, 朱璇, 热合满·艾拉. 果胶酶澄清和田红葡萄汁工艺研究[J]. 新疆农业科学,2010,47(6):1102−1106. [HALIHABAI AYMGL, ZHU X, AILA RHM. Pectinase clarification of hetian red grape juice[J]. Xinjiang Agricultural Sciences,2010,47(6):1102−1106. HALIHABAI AYMGL, ZHU X, AILA RHM. Pectinase clarification of hetian red grape juice[J]. Xinjiang Agricultural Sciences, 2010, 47(6): 1102 - 1106.