响应面法优化桔青霉PA-33菌株的发酵工艺
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摘要
【目的】优化桔青霉Penicillium citrinum PA-33发酵培养基和发酵条件,以提高桔青霉PA-33的抗菌活性。【方法】采用单因素试验确定桔青霉PA-33发酵所需最适基础培养基、碳氮源和无机盐,并利用响应面法设计确定最适发酵培养基配方;在发酵条件单因素试验基础上,采用三元二次通用旋转组合设计和频率分析法优化其最适发酵条件组合。【结果】经优化后,最佳发酵培养基配方为马铃薯汁液219.91 g·L-1、甘露醇34.11 g·L-1、黄豆粉6.25 g·L-1;最适发酵条件为装液量50 mL、接种量3.5%(φ)、发酵温度28℃,摇床转速150 r·min-1、发酵时间12 d。优化后发酵液对大肠埃希菌Escherichia coli的抑菌圈直径为28.99 mm,较优化前抑菌圈直径(18.73 mm)增加了10.26 mm。【结论】采用响应面法、三元二次通用旋转组合设计和频率分析法优化发酵工艺,显著提高了桔青霉PA-33发酵液的抗菌活性,为该菌株的抗菌活性物质的分离以及工业化生产提供依据。
【Objective】To optimize the medium composition and fermentation conditions of Penicillium citrinum PA-33 to improve its antibacterial activity. 【Method】Single factor experiments were used to determine the optimum basic medium, carbon sources, nitrogen sources and inorganic salts. The optimum formulation of fermentation medium was determined by response surface methodology. On the basis of single factor tests of fermentation conditions, the optimal combination of fermentation conditions was optimized by ternary quadratic rotation unitized design and frequency analysis method. 【Result】The optimal medium composition were: Potato juice 219.91 g·L-1, mannitol 34.11 g·L-1 and soybean powder 6.25 g·L-1. The optimal fermentation conditions were: Liquid volume 50 mL, inoculation concentration 3.5%(φ), culture temperature 28℃, rotation speed 150 r·min-1 and fermentation for 12 d. The inhibition zone diameter of fermentation broth after optimization on Escherichia coli reached 28.99 mm, increased by 10.26 mm compared with 18.73 mm under the original conditions. 【Conclusion】Response surface methodology, ternary quadratic rotation unitized design and frequency analysis method significantly enhance the antibacterial activity of fermentation broth of Penicillium citrinum PA-33, and this study provides a basis for isolation of antibacterial active substances and industrial production of this strain.
【Objective】To optimize the medium composition and fermentation conditions of Penicillium citrinum PA-33 to improve its antibacterial activity. 【Method】Single factor experiments were used to determine the optimum basic medium, carbon sources, nitrogen sources and inorganic salts. The optimum formulation of fermentation medium was determined by response surface methodology. On the basis of single factor tests of fermentation conditions, the optimal combination of fermentation conditions was optimized by ternary quadratic rotation unitized design and frequency analysis method. 【Result】The optimal medium composition were: Potato juice 219.91 g·L-1, mannitol 34.11 g·L-1 and soybean powder 6.25 g·L-1. The optimal fermentation conditions were: Liquid volume 50 mL, inoculation concentration 3.5%(φ), culture temperature 28℃, rotation speed 150 r·min-1 and fermentation for 12 d. The inhibition zone diameter of fermentation broth after optimization on Escherichia coli reached 28.99 mm, increased by 10.26 mm compared with 18.73 mm under the original conditions. 【Conclusion】Response surface methodology, ternary quadratic rotation unitized design and frequency analysis method significantly enhance the antibacterial activity of fermentation broth of Penicillium citrinum PA-33, and this study provides a basis for isolation of antibacterial active substances and industrial production of this strain.
引文
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[15]梁新乐,黄莹莹,张虹,等.响应面法优化桔青霉F-5-5核酸酶P1发酵培养基碳氮源[J].核农学报,2011,25(1):57-61.
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[19]喻晨,张亚雄,赵劼,等.响应面法优化桔青霉产核酸酶P1培养基[J].食品科学,2011,32(17):283-286.
[20]MENG L H,LIU Y,LI X M,et al.Citrifelins A and B,citrinin adducts with a tetracyclic framework from cocultures of marine-derived isolates of Penicillium citrinum and Beauveria felina[J].J Nat Prod,2015,78(9):2301-2305.
[2]郑婕施,江北,张琪,等.青霉属真菌活性代谢产物研究进展[J].工业微生物,2017(06):50-56.
[3]LU Z Y,LIN Z J,WANG W L,et al.Citrinin dimers from the halotolerant fungus Penicillium citrinum B-57[J].J Nat Prod,2008,71(4):543-546.
[4]DU L,LI D,ZHANG G,et al.Novel carbon-bridged citrinin dimers from a volcano ash-derived fungus Penicillium citrinum and their cytotoxic and cell cycle arrest activities[J].Tetrahedron,2010,66(47):9286-9290.
[5]CLARK B R,CAPON R J,LACEY E,et al.Citrinin revisited:From monomers to dimers and beyond[J].Org.Biomol Chem,2006,4(8):1520-1528.
[6]LIU H C,DU L,ZHU T J,et al.Two new citrinin di-mers from a volcano ash-derived fungus,Penicillium citrinum HGY1-5[J].Helv Chim Acta,2010,93(11):2224-2230.
[7]NI M,LIN W L,YANG P,et al.A novel citrinin derivative from the marine-source fungus Penicillium citrinum[J].Acta Pharm,2015,50(2):203-206.
[8]LIU Y,LI X M,MENG L H,et al.Bisthiodiketopiperazines and acorane sesquiterpenes produced by the marine-derived fungus Penicillium adametzioides AS-53 on different culture media[J].J Nat Prod,2015,78(6):1294-1299.
[9]申光辉,郑丽君,张志清,等.解淀粉芽胞杆菌PC2产抑菌物质培养基及发酵条件优化[J].微生物学通报,2017,44(6):1358-1369.
[10]王昊鹏,吴黎明,赵柳微,等.烟曲霉素发酵培养基的优化研究[J].食品工业科技,2017:1-14.
[11]RUQAYYAH T I D,JAMAL P,ALAM M Z,et al.Application of response surface methodology for protein enrichment of cassava peel as animal feed by the white-rot fungus Panus tigrinus M609RQY[J].Food Hydrocolloid,2014,42:298-303.
[12]MUTHUKUMAR V,RAJESH N,VENKATASAMY R,et al.Mathematical modeling for radial overcut on electrical discharge machining of incoloy 800 by response surface methodology[J].Procedia Materials Science,2014,6:1674-1682.
[13]YANG P,FANG M,LIU Y W.Optimization of a phase adjuster in a thermo-acoustic stirling engine using response surface methodology[J].Energy Procedia,2014,61:1772-1775.
[14]李莉,张赛,何强,等.响应面法在试验设计与优化中的应用[J].实验室研究与探索,2015,34(8):41-45.
[15]梁新乐,黄莹莹,张虹,等.响应面法优化桔青霉F-5-5核酸酶P1发酵培养基碳氮源[J].核农学报,2011,25(1):57-61.
[16]谢祥聪,刘琴英,蒋冬花,等.淡色生赤壳菌(Bionectriao chroleuca)Bo-1菌株产生抗菌物质的发酵条件优化[J].生物技术通报,2014(1):166-170.
[17]郭雷,朱文成,刘玮炜,等.抗菌活性海洋真菌HN4-13的鉴定及其发酵优化[J].微生物学通报,2013,40(6):951-958.
[18]胡一峰,曹一岚,黄俊龙.美伐他汀发酵菌种筛选和发酵条件优化[J].浙江化工,2012,43(2):4-6.
[19]喻晨,张亚雄,赵劼,等.响应面法优化桔青霉产核酸酶P1培养基[J].食品科学,2011,32(17):283-286.
[20]MENG L H,LIU Y,LI X M,et al.Citrifelins A and B,citrinin adducts with a tetracyclic framework from cocultures of marine-derived isolates of Penicillium citrinum and Beauveria felina[J].J Nat Prod,2015,78(9):2301-2305.