一株假丝酵母菌SF260生产槐糖脂培养基及发酵工艺的优化
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摘要
以高产槐糖脂假丝酵母菌(Candida bombicola)SF260为生产菌,利用单因素和正交试验优化槐糖脂的发酵培养基配方,并进一步优化其发酵工艺条件。结果表明,槐糖脂的最适发酵培养基配方:葡萄糖60 g/L,转基因大豆油80 g/L,酵母粉3 g/L,Na_2HPO_4·12H_2O 3g/L,KH_2PO_4 1g/L,槐糖脂产量达88.79 g/L,较优化前提高了27.22%。菌株SF260在5 L发酵罐内的最佳发酵工艺条件:在发酵液中葡萄糖、转基因大豆油质量浓度分别降至10 g/L后,采用连续流加补料的方式控制葡萄糖、转基因大豆油质量浓度为10～15 g/L,如果pH降至3.5以下,控制pH为3.5,转速400 r/min,发酵时间204 h。在此优化条件下,槐糖脂产量可达331.26 g/L,发酵强度达1.62 g/(L·h)。
Using Candida bombicola SF260 with high-yield sophorolipid as fermentation starter, the fermentation medium formula for sophorolipid production was optimized by single factor experiments and orthogonal tests, and the fermentation process conditions were further optimized. The results showed that the optimum fermentation medium formula for sophorolipid production by C. bombicola SF260 were as follows: glucose 60 g/L,transgenic soybean oil 80 g/L, yeast powder 3 g/L, Na_2HPO_4·12H_2O 3g/L, KH_2PO_4 1g/L. The fermentation yield of sophorolipid could reach 88.79 g/L,which increased by 27.22% compared with the original medium. The optimum fermentation process conditions of strain SF260 in 5 L bioreactor were as follows: after the concentration of glucose and transgenic soybean oil in the fermentation broth decreased to 10 g/L, the concentration of glucose and transgenic soybean oil was controlled to 10-15 g/L by continuous fed-batch fermentation. When the pH reduced to below 3.5, the pH value was controlled to 3.5, the rotational speed was 400 r/min. Under these optimal conditions, the yield of sophorolipids reached 331.26 g/L after 204 h fermentation and the fermentation intensity reached 1.62 g/(L·h).
Using Candida bombicola SF260 with high-yield sophorolipid as fermentation starter, the fermentation medium formula for sophorolipid production was optimized by single factor experiments and orthogonal tests, and the fermentation process conditions were further optimized. The results showed that the optimum fermentation medium formula for sophorolipid production by C. bombicola SF260 were as follows: glucose 60 g/L,transgenic soybean oil 80 g/L, yeast powder 3 g/L, Na_2HPO_4·12H_2O 3g/L, KH_2PO_4 1g/L. The fermentation yield of sophorolipid could reach 88.79 g/L,which increased by 27.22% compared with the original medium. The optimum fermentation process conditions of strain SF260 in 5 L bioreactor were as follows: after the concentration of glucose and transgenic soybean oil in the fermentation broth decreased to 10 g/L, the concentration of glucose and transgenic soybean oil was controlled to 10-15 g/L by continuous fed-batch fermentation. When the pH reduced to below 3.5, the pH value was controlled to 3.5, the rotational speed was 400 r/min. Under these optimal conditions, the yield of sophorolipids reached 331.26 g/L after 204 h fermentation and the fermentation intensity reached 1.62 g/(L·h).
引文
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[22]JIA X Q,QI L,ZHANG Y G,et al.Computational fluid dynamics simulation of a novel bioreactor for sophorolipid production[J].Chinese JChem Eng,2017,25(6):732-740.
[2]郝建安,张晓青,杨波,等.微生物表面活性剂应用新进展[J].生物技术,2017,27(4):396-402.
[3]伏圣秘.槐糖脂对作物病原真菌的抗菌作用及其新型生物杀菌剂的开发[D].济南:齐鲁工业大学,2014.
[4]ASHBY R D,SOLAIMAN D K Y,FOGLIA T A.Property control of sophorolipids:influence of fatty acid substrate and blending[J].Biotechnol Lett,2008,30(6):1093-1100.
[5]BANAT I M,FRANZETTI A,GANDOLFI I,et al.Microbial biosurfactants production,applications and future potential[J].Appl Microbiol Biot,2010,87(2):427-444.
[6]陈静,张云瑞,宋欣,等.槐糖脂的生产及其应用研究进展[J].食品科学,2007,28(8):525-529.
[7]刘青芝,郭学平,朱希强,等.槐糖脂的微生物合成及其应用研究进展[J].食品与药品,2009,11(11):51-55.
[8]ASMER H J,LANG S,WAGNER F,et al.Microbial production,Structure elucidation and bioconversion of sophorose lipids[J].J Am Oil Chem Soc,1988,65(9):1460-1466.
[9]李慧,马晓静,郭伟,等.槐糖脂的生物合成[J].安徽农业科学,2013,41(17):7425-7428.
[10]SHAO L,SONG X,MA X,et al.Bioactivities of sophorolipid with different structures against human esophageal cancer cells[J].J Surg Res,2012,173(2):286-291.
[11]李俊峰,门晋名,王裕宽,等.槐糖脂的生物合成及最新应用[J].应用化工,2013,42(2):363-366.
[12]HIRATA Y,RYU M,ODA Y,et al.Novel characteristics of sophorolipids,yeast glycolipid biosurfactants,as biodegradable low-foaming surfactants[J].J Biosci Bioeng,2009,108(2):142-146.
[13]KIM S Y,OH D K,LEE K H,et al.Effect of soybean oil and glucose on sophorose lipid fermentation by Torulopsis bombicola in continuous culture[J].Appl Microbiol Biot,1997,48(1):23-26.
[14]MARCHANT R,BANAT I M.Biosurfactants:A sustainable replacement for chemical surfactants[J].Biotechnol Lett,2012,34(9):1597-1605.
[15]VAN BOGAERT I N,SAERENS K,DE MUYNCK C,et al.Microbial production and application of sophorolipids[J].Appl Microbiol Biot,2007,76(1):23-43.
[16]MASAAKI K S,YUKA Y S,HORIUCHI J I.Efficient production of sophorolipids by Starmerella bombicola using a corncob hydrolysate medium[J].J Biosci Bioeng,2015,119(3):317-322.
[17]KIM Y B,YUN S H,KIM E K.Enhanced sophorolipid production by feeding-rate-controlled fed-batch culture[J].Bioresource Technol,2009,100(23):6028-6032.
[18]郭伟.槐糖脂的优化生产及其在石油工业的应用研究[D].济南:山东大学,2014.
[19]申靖.拟威克酵母利用中等碳链长度烷烃合成槐糖脂的研究[D].济南:山东大学,2012.
[20]翟秋倩.槐糖脂生物表面活性剂高产菌株构建和高效发酵工艺优化[D].青岛:青岛科技大学,2017.
[21]杨利,张旭,谭文松.溶氧水平与搅拌转速对发酵生产透明质酸分子量的影响[J].华东理工大学学报,2008,34(6):805-808.
[22]JIA X Q,QI L,ZHANG Y G,et al.Computational fluid dynamics simulation of a novel bioreactor for sophorolipid production[J].Chinese JChem Eng,2017,25(6):732-740.