混料设计及响应面法优化大黄药渣固态发酵生产漆酶的工艺
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摘要
通过优化大黄药渣固体培养基的组成及发酵条件,以期最大程度提高发酵生产漆酶的活力。采用D-最优混料设计对固体培养基碳源组成进行优化,优化后得到的固体培养基组成为:小麦麸皮47.2%,大黄药渣47.2%,葡萄糖1.6%,蛋白胨2.5%,氯化钙0.48%,磷酸二氢钾0.5%,硫酸铜0.02%,硫酸镁0.5%。通过中心组合设计响应面试验建立二次回归模型对固态发酵条件进行优化,确定最优发酵条件为发酵初始pH 4.3,发酵温度32.5℃,固液质量体积比1.47∶1,发酵时间7.5 d。在此条件下,发酵生产的漆酶酶活为7 602.1 IU/mL,与模型预测值相对误差在0.6%。
Solid fermentation medium formula for laccase with coriolus hirsutus and the fermentation conditions were optimized to improve laccase activity. The D-optimal design was firstly used to optimize components of carbon resources in the medium. The optimal medium compositions obtained were as follows: wheat barn 47.2%,residues of Chinese rhubarb 47.2%,glucose 1.6%,peptone 2.5%,CaCl20.48%,KH2 PO40.5%,CuSO40.02% and MgSO40.5%.The response surface experiment of center composite design(CCD) was applied to establish the quadratic regression model for optimization of solid fermentation conditions. And the optimal fermentation conditions were as follows: initial culture medium pH 4.3, fermentation temperature 32.5 ℃, solid-liquid ratio 1.47:1(w/V) and fermentation time 7.5 d.Under the optimum conditions, the activity of laccase reached 7 602.1 IU/mL, the relative error to the predictive value was 0.6%.
Solid fermentation medium formula for laccase with coriolus hirsutus and the fermentation conditions were optimized to improve laccase activity. The D-optimal design was firstly used to optimize components of carbon resources in the medium. The optimal medium compositions obtained were as follows: wheat barn 47.2%,residues of Chinese rhubarb 47.2%,glucose 1.6%,peptone 2.5%,CaCl20.48%,KH2 PO40.5%,CuSO40.02% and MgSO40.5%.The response surface experiment of center composite design(CCD) was applied to establish the quadratic regression model for optimization of solid fermentation conditions. And the optimal fermentation conditions were as follows: initial culture medium pH 4.3, fermentation temperature 32.5 ℃, solid-liquid ratio 1.47:1(w/V) and fermentation time 7.5 d.Under the optimum conditions, the activity of laccase reached 7 602.1 IU/mL, the relative error to the predictive value was 0.6%.
引文
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[3] CHAMPAGNE P P, RAMSAY J A. Dye decolorization and detoxification by laccase immobilized on porous glass beads[J]. Bioresource Technology, 2010,101(7):2230-2235. DOI:10.1016/j.biortech.2009.11.066.
[4] HSU C A, WEN T N, SU Y C, et al. Biological degradation of anthroquinone and azo dyes by a novel laccase from Lentinus sp.[J]. Environmental Sciences, 2012,46(9):5109-5117.DOI:10.1021/es2047014.
[5]李峰,王娜,张师愚,等.中药药渣的综合利用及其研究进展[J].中国医药工业杂志,2016,47(10):1322-1326. DOI:10.16522/j.cnki.cjph.2016.10.026.
[6] RANGABHASHIYAM S, SUGANYA E, SELVARAJU N,et al. Significance of exploiting non-living biomaterials for the biosorption of wastewater pollutants[J]. World Journal of Microbiology and Biotechnolog, 2014,30(6):1669-1689. DOI:10.1007/s11274-014-1599-y.
[7] YING Z, AMMAIYAPPAN S, JONATHAN W C W. Evaluation of humic substances during co-composting of food waste,sawdust and chinese medicinal herbal residues[J]. Bioresource Technology, 2014,168:229-234. DOI:10.1016/j.biortech.2014.05.070.
[8] WU Y R., LUO Z H, KWOK-KEI C R, et al. Purification and characterization of an extracellular laccase from the anthracene-degrading fungus Fusarium solani MAS2[J].Bioresource Technology,2010,101(24):9772-9777. DOI:10.1016/j.biortech.2010.07.091.
[9]柴新义,安双登,盛硕,等.产漆酶真菌筛选及其产酶条件的优化[J].西北农林科技大学学报(自然科学版),2015(3):205-210. DOI:10.13207/j.cnki.jnwafu.2015.03.021.
[10]凡军民,高大响,刘进华,等.树舌灵芝深层发酵培养基的优化研究[J].食品与机械,2010(4):23-25. DOI:10.3969/j.issn.1003-5788.2010.04.006.
[11] SHIN K S, LEE Y J. Purification and characterization of a new member of the laccase family from the white-rot basidiomycete coriolus hirsutus[J]. Archives of Biochemistry and Biophysics,2000,384(1):109-115. DOI:10.1006/abbi.2000.2083.
[12]林俊芳,刘志明,陈晓阳,等.真菌漆酶的酶活测定方法评价[J].生物加工过程,2009,7(4):1-8. DOI:10.3969/j.issn.1762-3678.2009.04.001.
[13]岳鹍,潘志恒,孙勇民.中药药渣发酵生产毛云芝菌漆酶培养基的工艺研究[J].食品与机械,2015,31(5):47-50. DOI:10.13652/j.issn.1003-5788.2015.05.011.
[14]孙瑞雪.食品级漆酶的发酵优化及分离纯化[D].天津:天津科技大学,2014.
[15]张永.杏鲍菇菌渣固态发酵产漆酶及其在染料脱色中的应用[D].无锡:江南大学,2017.