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首页> 外文期刊>Bioenergy research >Production, Purification, and Characterization of α-Amylase from Solventogenic Clostridium sp. BOH3
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Production, Purification, and Characterization of α-Amylase from Solventogenic Clostridium sp. BOH3

机译：溶剂型梭状芽孢杆菌α-淀粉酶的生产，纯化和表征硼酸3

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摘要

A solventogenic strain of Clostridium sp. BOH3 produces extracellular α-amylase (7.15 U/mg protein) in reinforced clostridial medium supplemented with sugarcane bagasse hydrolysate (1 % w/v) and a small amount of starch (0.1 % w/v), which is essential for the expression of α-amylase. In the presence of α-amylase, BOH3 utilizes starch directly without any pretreatment and produces butanol almost equivalent (∼90 %) to the production of butanol from glucose. α-Amylase can be purified from culture supernatant by using one-step weak anion exchange chromatography with a yield of 43 %. In peptide fingerprinting analysis, this enzyme shows homology with α-amylase produced by Clostridium acetobutylicum ATCC824. However, the molecular weight is 54 kDa, which is smaller than α-amylase of ATCC824 (84 kDa). This enzyme has optimum temperature at 45–50 °C and optimum pH at 4.5–5.5. Under this condition, the enzyme activity is 91.32 U/mg protein, and its Km and Vmax values are 1.71 ± 0.02 mg/ml and 96.13 ± 0.15 μmol/min/mg protein, respectively. Activity of this α-amylase can be enhanced (>1.5 times) by addition of Ca2+ and Co2+ and its activity can be maintained at an acidic pH (pH 3–5) for about 24 h. These unique characteristics suggest that this enzyme can be used for saccharification of starch for production of biofuel in one pot.
机译：产气荚膜梭菌的致溶剂菌株。 BOH3在增强的梭菌培养基中补充蔗糖蔗渣水解液（1％w / v）和少量淀粉（0.1％w / v）产生胞外α-淀粉酶（7.15U / mg蛋白）。 α-淀粉酶。在存在α-淀粉酶的情况下，BOH3无需任何预处理即可直接利用淀粉，并生成与葡萄糖生产的丁醇几乎相等的丁醇（〜90％）。可以使用一步弱阴离子交换色谱法从培养上清液中纯化α-淀粉酶，收率为43％。在肽指纹分析中，该酶显示与丙酮丁醇梭菌ATCC824产生的α-淀粉酶同源。但是，分子量为54 kDa，小于ATCC824的α-淀粉酶（84 kDa）。该酶的最佳温度为45–50°C，最佳pH为4.5–5.5。在此条件下，酶活性为91.32U / mg蛋白质，其Km和Vmax值分别为1.71±0.02 mg / ml和96.13±0.15μmol/ min / mg蛋白质。通过添加Ca2 +和Co2 +，可以增强该α-淀粉酶的活性（> 1.5倍），并且其活性可以在酸性pH（pH 3-5）下保持约24小时。这些独特的特征表明该酶可用于将淀粉糖化以在一锅中生产生物燃料。

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  • 来源
    《Bioenergy research》 |2014年第1期|132-141|共10页
  • 作者

    Gobinath Rajagopalan; Jianzhong He; Kun Lin Yang;

  • 作者单位

    1.Department of Chemical and Bimolecular Engineering National University of Singapore 4 Engineering Drive 4 Block E5-02-09 Singapore 117576 Singapore;

    2.Department of Civil and Environmental Engineering National University of Singapore 1 Engineering Drive 2 Block E1A-07-03 Singapore 117576 Singapore;

    1.Department of Chemical and Bimolecular Engineering National University of Singapore 4 Engineering Drive 4 Block E5-02-09 Singapore 117576 Singapore;

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  • 正文语种 eng
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  • 关键词

    α-Amylase ABE fermentation Solventogenic Clostridium sp. Starch utilization Maltose production;

    机译：α-淀粉酶ABE发酵溶剂型梭状芽孢杆菌淀粉利用麦芽糖生产;

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