嗜麦芽寡养单胞菌DHHJ固态发酵羽毛角蛋白的研究
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摘要
我国的羽毛资源极其丰富,羽毛中含有大量的角蛋白,角蛋白作为一种硬性蛋白,具有不溶于水和抗分解的性质,一般条件下不溶解,且不能被通常的蛋白酶水解,因此它们大部分没有被充分利用,有的甚至造成了环境污染。而近年来畜牧业及氨基酸工业的大力发展使得蛋白资源极其紧缺,因此研究最有效的处理方法使废弃的羽毛角蛋白直接转变成蛋白质或复合氨基酸,使之能直接得到应用或作为进一步提纯氨基酸的廉价易得的原料,从而变废为宝,既保护了环境又实现了资源利用,这将具有十分显著的经济效益和深远的社会效益。
羽毛废弃物的处理利用方法有很多,但这些方法都有缺点,不适宜大规模工业生产。近年来研究表明,固态发酵法具有节水、节能的独特优势,而且没有废液产生,是一种清洁生产技术,因此,当前应大力研究微生物固态发酵羽毛角蛋白工艺以及发酵产物的利用方式,使废弃的羽毛能得到高效利用,变废为宝,既保护了环境又有利于资源利用。
本课题针对羽毛角蛋白废弃物的资源化利用,进行了如下的研究：
首次将东华大学环境学院筛选、分离纯化,并经过诱变处理后的嗜麦芽寡养单胞菌DHHJ应用于固态发酵羽毛角蛋白工艺上,并对发酵产物进行了分析,得到了发酵产物的应用方向,同时进行了产物应用的经济性分析,研究结果如下：
1、羽毛固态发酵工艺可行性研究
在嗜麦芽寡养单胞菌液态发酵羽毛优化后的发酵条件下,通过改变种龄,接种量和发酵时间来对比菌株的液态发酵和固态发酵,实验结果表明在相同条件下,固态发酵羽毛工艺是可行的。
2、固态发酵羽毛角蛋白工艺的优化
经优化实验后得到嗜麦芽寡养单胞菌固态发酵羽毛的最佳工艺是：初始pH为7.5、培养温度为35℃、培养时间为4天、培养基组成是麸皮和干酪素,固液比是1：1.5,在该发酵条件下嗜麦芽寡养单胞菌的产酶能力及降解羽毛能力显著高于未优化发酵条件。其中,蛋白质含量达到30.72 mg/mL,粗酶液酶活力达到26.49 U/mL。这说明该菌株固态发酵羽毛角蛋白具有潜在的实际开发价值。
3、发酵产物分析
通过分析发酵产物,得出羽毛发酵液中氨基酸和微量元素的组分分布,以及可溶蛋白分子量分布。根据发酵液组成特点,认为合适的应用领域是通过表面吸收方式利用,如洗护用品或肥料。
4、两种发酵方法的经济性分析和比较
通过对固态发酵产物生产过程的成本分析,并与液态发酵条件的情况对比可知,在生产过程中,得到相同质量的发酵产物,固态发酵条件下的成本低于液态发酵条件下。同时,对于发酵产物中残留的羽毛粉,建议部分代替鱼粉或作为辅料直接加入鱼粉中,能有效的提高饲料产品质量,同时降低饲料成本。
There are rich and huge quantity feather resources in China.However, most of them are not well utilized, left idle and even becoming pollution source. Contrast to normal feather processing methods, microorganism method has its prominat advantage; most present ways still hold on strain isolation and keratinase purification, but not expand and not evolve the zymotechnique into industrlization. It's even hard to find the documentation aboutthe application of feather fermentation products.
There are many methods of using feather waste, but all of the methods have shortcoming. In recently years, research shows solid-state fermentations (SSF) is a kind of cleaner production technologies, this method with the unique superiority of water and energy saving, and no liquid produced. In this paper, we would take the hight priority to study how to amplify the zymotechnique of feather that uses microorganism method, to make good use of abandoned feather by which protects the environment from being polluted. On the other hand, the aim is on changing abandoned feather into resource.
It's the first time to record that scaling zymotechnique of Stenotrophomonas maltophilia DHHJ that is selected, isolated, purified and mutagenize by Envrionment Institute of Donghua University. This method has remarkable effect on fertilization in leaf surface and hair caring.
Firstly, Feather Zymotechnique under SSF
In order to industrialize the utilization of waste feather by microbiological method, the solid-state fermentation feather feasibility test and research for Stenotrophomonas maltophilia DHHJ based on the study of fermentation in Erlenmeyer flask was investigated. Further, this paper analyzed the fermentation process and optimized the technological parameters. Result shows that solid-state fermentation feather is feasible, and provided the reliable basis for the use of the feather waste.
Secondly, mmobilization Optimization of Fermentation Conditions
In order to industrialize the utilization of waste feather by microbiological method, the test of solid-state fermentation for feather with Stenotrophomonas maltophilia DHHJ was studied based on its liquid-state fermentation previously. The results showed that the optimal solid state fermentation was initial pH 7.5 at temperature 35℃for four days, the medium was composed of bran and casein, and the solid-liquid ratio was 1:1.5. Under the optimal conditions, the abilities of keratinase production and feather degradation of strain DHHJ were much better than those under the liquid-state fermentation, the protein content was 30.72 mg/mL and the activity of keratinase was 26.49 U/mL.
Thirdly, Fermentation Products Analysis
It's to get the proportion of aminophenol, microelement, and soluable protetin molecules. Comparing to existed experment reports, it may conclude the enzyme system related to bacteria strain DHHJ and feathers. Inhence, it is conferred that the industry fields of the application can be cleaning products and fertilizers.
Lastly, Cost Analysis and comparison between SSF and LSF
Through the solid-state fermentation product process cost analysis, and with liquid fermentation conditions contrast knowable, we can know in the process of production, getting the same quality of fermented product, solid-state fermentation conditions costing less than liquid fermentation conditions. Economically analysis, raw material price is relatively low, therefore has the considerable economical development value.
羽毛废弃物的处理利用方法有很多,但这些方法都有缺点,不适宜大规模工业生产。近年来研究表明,固态发酵法具有节水、节能的独特优势,而且没有废液产生,是一种清洁生产技术,因此,当前应大力研究微生物固态发酵羽毛角蛋白工艺以及发酵产物的利用方式,使废弃的羽毛能得到高效利用,变废为宝,既保护了环境又有利于资源利用。
本课题针对羽毛角蛋白废弃物的资源化利用,进行了如下的研究：
首次将东华大学环境学院筛选、分离纯化,并经过诱变处理后的嗜麦芽寡养单胞菌DHHJ应用于固态发酵羽毛角蛋白工艺上,并对发酵产物进行了分析,得到了发酵产物的应用方向,同时进行了产物应用的经济性分析,研究结果如下：
1、羽毛固态发酵工艺可行性研究
在嗜麦芽寡养单胞菌液态发酵羽毛优化后的发酵条件下,通过改变种龄,接种量和发酵时间来对比菌株的液态发酵和固态发酵,实验结果表明在相同条件下,固态发酵羽毛工艺是可行的。
2、固态发酵羽毛角蛋白工艺的优化
经优化实验后得到嗜麦芽寡养单胞菌固态发酵羽毛的最佳工艺是：初始pH为7.5、培养温度为35℃、培养时间为4天、培养基组成是麸皮和干酪素,固液比是1：1.5,在该发酵条件下嗜麦芽寡养单胞菌的产酶能力及降解羽毛能力显著高于未优化发酵条件。其中,蛋白质含量达到30.72 mg/mL,粗酶液酶活力达到26.49 U/mL。这说明该菌株固态发酵羽毛角蛋白具有潜在的实际开发价值。
3、发酵产物分析
通过分析发酵产物,得出羽毛发酵液中氨基酸和微量元素的组分分布,以及可溶蛋白分子量分布。根据发酵液组成特点,认为合适的应用领域是通过表面吸收方式利用,如洗护用品或肥料。
4、两种发酵方法的经济性分析和比较
通过对固态发酵产物生产过程的成本分析,并与液态发酵条件的情况对比可知,在生产过程中,得到相同质量的发酵产物,固态发酵条件下的成本低于液态发酵条件下。同时,对于发酵产物中残留的羽毛粉,建议部分代替鱼粉或作为辅料直接加入鱼粉中,能有效的提高饲料产品质量,同时降低饲料成本。
There are rich and huge quantity feather resources in China.However, most of them are not well utilized, left idle and even becoming pollution source. Contrast to normal feather processing methods, microorganism method has its prominat advantage; most present ways still hold on strain isolation and keratinase purification, but not expand and not evolve the zymotechnique into industrlization. It's even hard to find the documentation aboutthe application of feather fermentation products.
There are many methods of using feather waste, but all of the methods have shortcoming. In recently years, research shows solid-state fermentations (SSF) is a kind of cleaner production technologies, this method with the unique superiority of water and energy saving, and no liquid produced. In this paper, we would take the hight priority to study how to amplify the zymotechnique of feather that uses microorganism method, to make good use of abandoned feather by which protects the environment from being polluted. On the other hand, the aim is on changing abandoned feather into resource.
It's the first time to record that scaling zymotechnique of Stenotrophomonas maltophilia DHHJ that is selected, isolated, purified and mutagenize by Envrionment Institute of Donghua University. This method has remarkable effect on fertilization in leaf surface and hair caring.
Firstly, Feather Zymotechnique under SSF
In order to industrialize the utilization of waste feather by microbiological method, the solid-state fermentation feather feasibility test and research for Stenotrophomonas maltophilia DHHJ based on the study of fermentation in Erlenmeyer flask was investigated. Further, this paper analyzed the fermentation process and optimized the technological parameters. Result shows that solid-state fermentation feather is feasible, and provided the reliable basis for the use of the feather waste.
Secondly, mmobilization Optimization of Fermentation Conditions
In order to industrialize the utilization of waste feather by microbiological method, the test of solid-state fermentation for feather with Stenotrophomonas maltophilia DHHJ was studied based on its liquid-state fermentation previously. The results showed that the optimal solid state fermentation was initial pH 7.5 at temperature 35℃for four days, the medium was composed of bran and casein, and the solid-liquid ratio was 1:1.5. Under the optimal conditions, the abilities of keratinase production and feather degradation of strain DHHJ were much better than those under the liquid-state fermentation, the protein content was 30.72 mg/mL and the activity of keratinase was 26.49 U/mL.
Thirdly, Fermentation Products Analysis
It's to get the proportion of aminophenol, microelement, and soluable protetin molecules. Comparing to existed experment reports, it may conclude the enzyme system related to bacteria strain DHHJ and feathers. Inhence, it is conferred that the industry fields of the application can be cleaning products and fertilizers.
Lastly, Cost Analysis and comparison between SSF and LSF
Through the solid-state fermentation product process cost analysis, and with liquid fermentation conditions contrast knowable, we can know in the process of production, getting the same quality of fermented product, solid-state fermentation conditions costing less than liquid fermentation conditions. Economically analysis, raw material price is relatively low, therefore has the considerable economical development value.
引文
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