黑曲霉HG-1固态发酵苹果渣和棉粕生产果胶酶的工艺研究
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摘要
果胶酶(pectinase)是世界四大酶制剂之一,是分解果胶质酶类的总称,在食品、酿酒、环保、医药、纺织及洗涤剂等行业应用十分广泛。果胶酶通常包括聚半乳糖醛酸酶(PG)、果胶质酶(PE)、果胶裂解酶(PL)等主要成分。不同组分的作用机理和酶学特性存在差异。黑曲霉是果胶酶工业化生产的主要菌种。为了进一步开发黑曲霉产果胶酶酶能力,本论文研究了以黑曲霉HG-1固态发酵苹果渣和棉粕生产果胶酶的工艺研究。
本文是采用价格低廉的农业废弃物苹果渣和棉粕为主要原料生产果胶酶,优化其生产工艺,并对果胶酶的部分酶学性质进行研究。研究中,以黑曲霉HG-1为生产菌种,采用单因子试验和正交试验进行固态发酵。试验结果表明,培养基为苹果渣10g、棉粕10g、(NH4)2SO4 0.2g、K2HPO4 0.06g、初始水分含量60%,及最适发酵条件为装料量20g干料/250ml三角瓶、30℃恒温培养48h时,所得果胶酶酶活力可达22248 U/g。
果胶酶酶促反应最适温度为45℃,最适pH为5.0;在50℃以下,pH 3.0～6.0时稳定性良好;Ca2+、Mg2+、Fe2+对该酶有激活作用,而Ba2+、Mn2+、Zn2+有抑制作用。
结果表明苹果渣和棉粕代替麸皮作为黑曲霉HG-1固态发酵生产果胶酶的主要原料在技术上具有可行性,还可以部分解决苹果渣和棉粕的综合利用问题。
Pectinase is one of the fourth of the Enzyme Preparations in the world and is collective call for any enzyme which breaks down pectic substance and pectinase is widely used in Foods, Medicine, Environmental Protection, Textile, Brewing, Detergent field.Primarily, it sontains components such as polygalacturonase, pectinesterase and pectinlyase. Each component differs from others in the mechanism of reaction and its characteristics.Aspergillus niger was the major genera of microorganisms producing the pectinase in industry. In order to exploit the capacity of Aspergillus niger in producing the enzyme, it is studying on process of producting pectinase from apple pomace and cottonseed meal by Aspergillus niger HG-1 in solid state fermentation in this paper.
It is on optimizing the culture medium and fermentation conditions of Aspergillus niger HG-1 to produce pectinase from apple pomace and cottonseed meal in solid state fermentation, and to reveal some prime characteristic of the pectinase in this paper. In this study, Monofactorial and orthogonal experiments were adopted in solid state fermentation. The results show that the activity of pectinase reached 22248 U/g when Aspergillus niger HG-1 was cultured at 30℃for 48h in the optimized medium containing 10g apple pomace,10g cottonseed meal,0.2g (NH4)2SO4 and 0.06g KH2PO4 at the optimized conditions including moisture content of solid substrate 60% (w/w) and 20g dry substrate /250ml flask.
The optimal temperature and pH of pectinase reaction were at 45℃and pH5.0. Pectinase was stable below 50℃and between pH3.0 and pH6.0. Ca2+,Mg2+ and Fe2+ increased pectinase activity whereas Ba2+,Mn2+,Zn2+ obviously inhibited it.
The results showed that it was technically and economically feasible to produce pectinase from apple pomace by Aspergillus niger HG-1 in solid statefermentation. It was also an effective way to make the best of apple pomace,an agricultural waste produced in larger quantities in China.
本文是采用价格低廉的农业废弃物苹果渣和棉粕为主要原料生产果胶酶,优化其生产工艺,并对果胶酶的部分酶学性质进行研究。研究中,以黑曲霉HG-1为生产菌种,采用单因子试验和正交试验进行固态发酵。试验结果表明,培养基为苹果渣10g、棉粕10g、(NH4)2SO4 0.2g、K2HPO4 0.06g、初始水分含量60%,及最适发酵条件为装料量20g干料/250ml三角瓶、30℃恒温培养48h时,所得果胶酶酶活力可达22248 U/g。
果胶酶酶促反应最适温度为45℃,最适pH为5.0;在50℃以下,pH 3.0～6.0时稳定性良好;Ca2+、Mg2+、Fe2+对该酶有激活作用,而Ba2+、Mn2+、Zn2+有抑制作用。
结果表明苹果渣和棉粕代替麸皮作为黑曲霉HG-1固态发酵生产果胶酶的主要原料在技术上具有可行性,还可以部分解决苹果渣和棉粕的综合利用问题。
Pectinase is one of the fourth of the Enzyme Preparations in the world and is collective call for any enzyme which breaks down pectic substance and pectinase is widely used in Foods, Medicine, Environmental Protection, Textile, Brewing, Detergent field.Primarily, it sontains components such as polygalacturonase, pectinesterase and pectinlyase. Each component differs from others in the mechanism of reaction and its characteristics.Aspergillus niger was the major genera of microorganisms producing the pectinase in industry. In order to exploit the capacity of Aspergillus niger in producing the enzyme, it is studying on process of producting pectinase from apple pomace and cottonseed meal by Aspergillus niger HG-1 in solid state fermentation in this paper.
It is on optimizing the culture medium and fermentation conditions of Aspergillus niger HG-1 to produce pectinase from apple pomace and cottonseed meal in solid state fermentation, and to reveal some prime characteristic of the pectinase in this paper. In this study, Monofactorial and orthogonal experiments were adopted in solid state fermentation. The results show that the activity of pectinase reached 22248 U/g when Aspergillus niger HG-1 was cultured at 30℃for 48h in the optimized medium containing 10g apple pomace,10g cottonseed meal,0.2g (NH4)2SO4 and 0.06g KH2PO4 at the optimized conditions including moisture content of solid substrate 60% (w/w) and 20g dry substrate /250ml flask.
The optimal temperature and pH of pectinase reaction were at 45℃and pH5.0. Pectinase was stable below 50℃and between pH3.0 and pH6.0. Ca2+,Mg2+ and Fe2+ increased pectinase activity whereas Ba2+,Mn2+,Zn2+ obviously inhibited it.
The results showed that it was technically and economically feasible to produce pectinase from apple pomace by Aspergillus niger HG-1 in solid statefermentation. It was also an effective way to make the best of apple pomace,an agricultural waste produced in larger quantities in China.
引文
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