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利用响应面法优化新分离的嗜热枯草芽孢杆菌K-18产羧甲基纤维素酶用于糖化反应

Carboxymethyl cellulase production optimization from newly isolated thermophilic Bacillus subtilis K-18 for saccharification using response surface methodology.

作者信息

Irfan Muhammad, Mushtaq Qudsia, Tabssum Fouzia, Shakir Hafiz Abdullah, Qazi Javed Iqbal

机构信息

Department of Biotechnology, University of Sargodha, University Road, Sargodha, 40100, Pakistan.

Microbial Biotechnology Laboratory, Department of Zoology, University of the Punjab, New Campus, Lahore, 54590, Pakistan.

出版信息

AMB Express. 2017 Dec;7(1):29. doi: 10.1186/s13568-017-0331-3. Epub 2017 Jan 31.

Abstract

In this study, a novel thermophilic strain was isolated from soil and used for cellulase production in submerged fermentation using potato peel as sole carbon source. The bacterium was identified by 16S rRNA gene sequencing technology. Central composite design was applied for enhanced production using substrate concentration, inoculum size, yeast extract and pH as dependent variables. Highest enzyme titer of 3.50 ± 0.11 IU/ml was obtained at 2% substrate concentration, 2% inoculum size, 1% yeast extract, pH 5.0, incubation temperature of 50 °C for 24 h of fermentation period. The crude enzyme was characterized having optimum pH and temperature of 7.0 and 50 °C, respectively. The efficiency of enzyme was checked by enzymatic hydrolysis of acid/alkali treated pine needles which revealed that 54.389% saccharification was observed in acid treated pine needles. These results indicated that the cellulase produced by the Bacillus subtilis K-18 (KX881940) could be effectively used for industrial processes particularly for bioethanol production.

摘要

在本研究中,从土壤中分离出一种新型嗜热菌株,并以马铃薯皮作为唯一碳源,用于深层发酵生产纤维素酶。采用16S rRNA基因测序技术对该细菌进行鉴定。以底物浓度、接种量、酵母提取物和pH作为自变量,应用中心复合设计提高产量。在底物浓度2%、接种量2%、酵母提取物1%、pH 5.0、培养温度50℃、发酵24小时的条件下,获得了最高酶活3.50±0.11 IU/ml。对粗酶进行表征,其最适pH和温度分别为7.0和50℃。通过对酸/碱处理的松针进行酶水解来检测酶的效率,结果表明酸处理的松针糖化率为54.389%。这些结果表明,枯草芽孢杆菌K-18(KX881940)产生的纤维素酶可有效地用于工业生产过程,特别是生物乙醇生产。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9444/5302012/b694cb89b85f/13568_2017_331_Fig1_HTML.jpg

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