产β-葡萄糖苷酶芽孢杆菌的筛选及水解大豆异黄酮糖苷研究

doi:10.3969/j.issn.1000-1565.2017.06.010

摘要/Abstract

摘要： 为获得能高效水解大豆异黄酮糖苷的芽孢杆菌,利用七叶苷平板分离法从动物的新鲜粪便中筛选产β-葡萄糖苷酶的芽孢杆菌菌株,然后对酶活最高的菌株进行种属鉴定和水解大豆异黄酮糖苷的研究.结果显示:筛选到1株酶活力较高的芽孢杆菌R2-2,经鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens);该菌株有较高的糖苷水解能力,黄豆苷水解率最高达到53.65%,染料木苷为48.80%.

关键词: β-葡萄糖苷酶, 芽孢杆菌, 筛选, 大豆异黄酮糖苷

Abstract: In order to obtain Bacillus strains capable of efficiently hydrolyze soybean isoflavone glycosides,this study screened Bacillus strains producing β-glucosidase from fresh feces of animals with the help of Aesculin separation culture medium.Then the strains with relatively higher β-glucosidase activity were identified and the ability of hydrolyzing isoflavone glucosides were studied.The results showed that a strains of Bacillus R2-2 with relatively higher β-glucosidase activity was screened out.The identity of strains R2-2 was Bacillus amyloliquefaciens.Strain R2-2 showed rather high efficiency in hydrolyzing isoflavone glucosides.The daidzin hydrolysis rate reached 53.65%,and genistin hydrolysis rate was 48.80%.

Key words: β-glucosidase, Bacillus, screen, soybean isoflavone glycosides

中图分类号:

S816

刘宏丽,郭晓军,狄聪颖, 郭威,朱宝成. 产β-葡萄糖苷酶芽孢杆菌的筛选及水解大豆异黄酮糖苷研究[J]. 河北大学学报(自然科学版), 2017, 37(6): 621-629.

LIU Hongli,GUO Xiaojun,DI Congying,GUO Wei,ZHU Baocheng. Screening of Bacillus strains producing β-glucosidase and study on hydrolysis of soybean isoflavone glycosides[J]. Journal of Hebei University (Natural Science Edition), 2017, 37(6): 621-629.

参考文献

[1] 李万林,钟姣姣,王少龙,等.大豆异黄酮生理功能及其检测方法的研究进展[J].饮料工业,2014,17(4):54-59. LI W L,ZHONG J J,WANG S L,et al.Research progresses in physiological functions of and detection methods for soybean isoflavones[J].Beverage Industry,2014,17(4):54-59.
[2] 何恩铭,沈瑞池,林志楷,等.大豆异黄酮抗肿瘤效应研究进展[J].亚热带植物科学,2013,42(4):356-360.DOI: 10.3969/j.issn.1009-7791.2013.04.019. HE E M,SHEN R C,LIN Z K,et al.A review on anti-tumor effects of soy isoflavones[J].Subtropical Plant Science,2013,42(4):356-360.DOI: 10.3969/j.issn.1009-7791.2013.04.019.
[3] 罗佳捷,范觉鑫,张彬.大豆异黄酮在动物生产中的研究进展[J].饲料博览,2012,1:9-11.DOI: 10.3969/j.issn.1001-0084.2012.01.004. LUO J J,FAN J X,ZHANG B.Research progress of soybean isoflavone in animal production[J].Feed Review,2012,1:9-11.DOI: 10.3969/j.issn.1001-0084.2012.01.004.
[4] KUMAR N B,CANTOR A,KATHY A R D,et al.The specific role of isoflavones on estrogen metabolism in premenopausal women[J].Cancer,2002,94(4): 1166 -1174.DOI: 10.1002/cncr.10320.
[5] 金凤燮.天然产物生物转化[M].北京:化学工业出版社,2009:223 -243.
[6] ROUTLEDGE E J,PARKER J,ODUM J,et al.Some alkyl hydroxy benzoate preservatives(parabens)are estrogenic [J].Toxicology and Applied Pharmacology,1998,153(1): 12-19.
[7] 杨晓宽.β-葡萄糖苷酶研究进展[J].河北科技师范学院学报,2012,26(1):77-81.DOI: 10.3969 / J.issn.1672-7983.2012.01.016. YANG X K.Research progress on β-glucosidase[J].Journal of Hebei Normal University of Science & Technology,2012,26(1):77-81.DOI: 10.3969 / J.ISSN.1672-7983.2012.01.016.
[8] HESSLER P E,LARSEN P E,CONSTANTINOU A I,et al.Isolation of isoflavones from soy-based fermentations of the erythromycin-producing bacterium Saccharopolyspora erythraea[J].Appl Microbiol Biotechnol,1997,47: 398-404.DOI: 10.1007/s002530050947.
[9] TSANGALIS D,ASHTON J F,MCGILL A E J,et al.Enzymatic transformation of isoflavone phytoestrogens in soy milk by β-glucosidase-producing Bifidobacteria[J].J Food Sci,2002,67:3104-3113.DOI: 10.1111/j.1365-2621.2002.tb08866.x.
[10] 潘艳,张益波,郭志敏,等.β-葡萄糖苷酶高产菌株筛选、诱变及其发酵的研究[J].食品研究与开发,2012,33(5):149-152.DOI:10.3969/j.issn.1005-6521.2012.05.043. PAN Y,ZHANG Y B,GUO Z M,et al.Study on screening and mutation of β-Glucosidase-producing strain[J].Food Research and Development,2012,33(5):149-152.DOI: 10.3969/j.issn.1005-6521.2012.05.043.
[11] 黄琴,朱婷,蒋承建,等.产β-葡萄糖苷酶的菌种的筛选鉴定及其酶学特性[J].基因组学与应用生物学,2011,30(5):590-595. DOI: 10.3969/gab.030.000590. HUANG Q,ZHU T,JIANG C J,et al.Isolation,identification and the enzymatic characteristics of the strain producing β-glucosidase[J].Genomics and Applied Biology,2011,30(5):590-595.DOI: 10.3969/gab.030.000590.
[12] 施特尔马赫.酶的测定方法[M].钱嘉渊,译.北京:中国轻工业出版社出版,1992:102-119.
[13] 沈萍,陈向东.微生物学实验[M].第4版.北京:高等教育出版社,2007.
[14] 杨继业,郭晓军,郭威,等.产有机酸芽孢杆菌的筛选、鉴定及产芽孢条件优化[J].饲料工业,2015,369(4):44-51.DOI:10.13302/j.cnki.fi.2015.04.010. YANG J Y,GUO X J,GUO W,et al.Screening and identification of a strain of Bacillus sp.producing organic acids and optimization of its spore production conditions[J].Feed Industry,2015,369(4):44-51.DOI:10.13302/j.cnki.fi.2015.04.010.
[15] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[16] 齐斌,刘贤金.产大豆异黄酮β-葡萄糖苷酶菌株的筛选及酶学性质研究[J].食品科学,2007,28(8): 290-293.DOI: 10.3321/j.issn:1002-6630.2007.08.069. QI B,LIU X J.Study on screen of soybean isoflavone β-glucosidase-producing strain and its enzymatic properties[J].Food Science,2007,28(8): 290-293.DOI: 10.3321/j.issn:1002-6630.2007.08.069.
[17] 潘利华,罗建平,罗水忠,等.高活性大豆异黄酮糖苷酶的分离原料筛选、纯化及性质研究[J].食品科学,2009,30(1):164-168.DOI:10.3321/j.issn:1002-6630.2009.01.039. PAN L H,LUO J P,LUO S Z,et al.Seleetion of plant source used for separating isoflavone-glucosidase with high aetivity and its purification and properties[J].Food Science,2009,30(1):164-168.DOI:10.3321/j.issn:1002-6630.2009.01.039.
[18] 高荣海,刘长江,李长彪,等.大豆异黄酮糖苷水解研究进展[J].粮食与油脂,2006(8): 10-12.DOI: 10.3969/j.issn.1008-9578.2006.08.003. GAO R H,LIU C J,Ll C B,et al.Research advance on hydrolyzation of soybean isonavone glycoside[J].Cereals & Oils,2006(8): 10-12.DOI:10.3969/j.issn.1008-9578.2006.08.003.
[19] 崔洪斌.大豆生物活性物质的开发与应用[M].北京:中国轻工业出版社,2001:178-179.
[20] 于卓腾.大豆黄酮对仔猪肠道微生物的影响和雌马酚产生菌的分离及其特性的研究[D].南京农业大学,2007. YU Z T.Effects of daidzein on the gastrointestinal mcicrobial community of piglets and isolation and study characterisation of equol-producing bacteria[D].Nanjing Agricultural University,2007.
[21] 杨守凤,徐建雄.β-葡萄糖苷酶转化大豆异黄酮及其保健功能的研究进展[J].饲料研究,2014,01:6-12.
[22] 刘德海,马焕,谢复红,等.一株产β-葡萄糖苷酶烟曲霉菌株的鉴定及其产酶特性[J].中国酿造,2015,34(6):118-122.DOI:10.11882/j.issn.0254-5071.2015.06.026. LIU D H,MA H,XIE F H,et al.Identification of β-glucosidase producing Aspergillus fumigatus and its enzyme production characteristics[J].China Brewing,2015,34(6):118-122.DOI:10.11882/j.issn.0254-5071.2015.06.026.
[23] 朱婧,覃拥灵,陈桂光,等.β-葡萄糖苷酶高产菌株的选育及酶法转化葡萄糖生产龙胆低聚糖[J].食品与发酵工业,2010,36(4):21-25.DOI:10.13995/j.cnki.11-1802/ts.2010.04.024. ZHU J,QIN Y L,CHEN G G,et al.Selection of the high β-glucosidase mutant and production of gentiooligosaccharid[J].Food and Fermentation Industries,2010,36(4):21-25.DOI:10.13995/j.cnki.11-1802/ts.2010.04.024.
[24] 张秋卓,蔡伟民.纤维素酶高产菌株的复合交替诱变选育[J].工业微生物,2008,38(6):32-37.DOI:10.3969/j.issn.1001-6678.2008.06.007. ZHANG Q Z,CAI W M.Selecuion of high cellulase-yielding mutant by compound mutagenesis[J].Industrial Microbiology,2008,38(6):32-37.DOI:10.3969/j.issn.1001-6678.2008.06.007.
[25] KRISHNA S H,PRASANTHI K,CHOWDARY G V.Simultaneous saccharification and fermentation of pretreated sugar cane leaves to ethanol[J].Process Biochemistry,1998,33(8):825-830.DOI.org/10.1016/S0032-9592(98)00051-X.
[26] RICCARDO N B,GIOVANNI S,ROSA P,et al.Selection,characterization and compareson of β-glucosidase from mould and yeasts employable for enological application[J].Enzyme and Microbial Technology,2004,35:58-66.
[27] 蒋文鸿.昌黎与济源产区高产β-葡萄糖苷酶野生非酿酒酵母的筛选及鉴定[D].杨凌:西北农林科技大学,2014. JIANG W H.Screening and identification of high producing β-glucosidase wild non-saccharomyces Yeast strain from JIYUAN and CHANGLI[D].Yangling: Northwest A & F University,2014.
[28] RAIMONDI S,RONCAGLIA L,LUCIA M D,et al.Bioconversion of soy isoflavones daidzin and daidzein by Bifidobacterium strains[J].Appl Microbiol Biotechnol,2009,81:943-950.DOI:1007/s00253-008-1719-4.
[29] SUBROTA HATI,SHILPA VIJ,BRIJ PAL SINGH.et al.β- Glucosidase activity and bioconversion of isoflavones during fermentation of soymilk[J].J Sci Food Agric,2015,95: 216-220.DOI:10.1002/jsfa.6743.
[30] OTIENO D O,ASHTON J F,SHAH N P.Evaluation of enzymic potential for biotransformation of isoflavone phytoestrogen in soymilk by Bifidobacterium animalis,Lactobacillus acidophilus and Lactobacillus casei [J].Food Research International,2006,39(4): 394-407.DOI.:org/10.1016/j.foodres.2005.08.010.
[31] BAU' T R,GARCIA S,IDA E I.Changes in soymilk during fermentation with kefir culture: oligosaccharides hydrolysis and isoflavone aglycone production[J].Food Sci Nutr,2015,66(8): 845-850.DOI:10.3109/09637486.2015.1095861.
[32] 陈红漫,赵璐,杨佳影,等.一株产胞外中性β-葡萄糖苷酶菌株的鉴定及酶学特性研究[J].食品科学,2009,30(15):160-163.DOI:10.3321/j.issn:1002-6630.2009.15.036. CHEN H M,ZHAO L,YANG J Y,et al.Identification of a novel Bacillus strain capable of producing neutral β-G lucosidase and characterization of its enzy matic properties[J].Food Science,2009,30(15): 160-163.DOI:10.3321/j.issn:1002-6630.2009.15.036.
[33] 王彩肖,武伟伟,李艳.产β-D-葡萄糖苷酶酵母菌的筛选及产酶性质研究[J].酿酒科技,2014,10:14-18.DOI:10.13746/j.njkj.2014.0167. WANG C X,WU W W,LI Y.Screening of yeast strains with high-yield of β-D-glucosidase and study on their β-D-glucosidase-producing properties[J].Liq-Mak Sci Technol,2014,10:14-18.DOI:10.13746/j.njkj.2014.0167.