CN111304185A - 一种纯固态发酵的低温淀粉酶的生产方法 - Google Patents
一种纯固态发酵的低温淀粉酶的生产方法 Download PDFInfo
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Abstract
本发明公开了一种纯固态发酵的低温淀粉酶的生产方法,配方包括:产低温淀粉酶为主,包含蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系的一种真菌(如黑曲霉菌或米曲霉菌等)或多种真菌组合,步骤一,菌种选取;步骤二,菌种培养;步骤三,培养基选取及称量;步骤四,培养基发酵处理;该发明,能够保证菌株快速生长繁殖快,代谢目的产物高效获得,代谢产物量高,综合活性比现有单一液体发酵技术生产的饲用低温淀粉酶提高5倍以上,能够弥补动物消化系统内源酶(淀粉酶)分泌的不足,显著提高动物饲粮中重要的能量来源(淀粉)的消化利用率,帮助满足动物生长性能的能量需求。
Description
技术领域
本发明涉及酶的生产、畜牧养殖技术领域,具体为一种纯固态发酵的低温淀粉酶的生产方法。
背景技术
近年来,随着我国人民的生活水平的不断提高,我国的饲料业、养殖业得到迅速发展,饲料资源不足已经成为阻碍我国饲料工业发展的一个重要原因。我国各地主要粮食作物差异较大,外源酶制剂的添加对潜在饲料资源的利用和新饲料资源的开发有较大的帮助。另一方面,随着人们生活水平的提高及环境意识的增强,政府也正积极通过禁止在饲料中使用抗生素、激素等方式来保障饲料和食品的安全,维护生态平衡,实现健康可持续的养殖模式。因此设计一种纯固态发酵的低温淀粉酶的生产方法,充分应用其产生的淀粉酶酶系综合活性强,酶系针对性强等特点全面提升动物饲粮主要能量来源(淀粉)的消化利用率,有利于弥补动物消化系统内源酶分泌不足,提升畜禽生产性能和新饲粮的开发利用。
发明内容
本发明的目的在于提供一种纯固态发酵的低温淀粉酶的生产方法,以解决上述背景技术中提出的问题。
为了解决上述技术问题,本发明提供如下技术方案:一种纯固态发酵的低温淀粉的生产方法,配方包括:产低温淀粉酶为主,包含蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系的单一真菌(如黑曲霉菌或米曲霉菌等)或多种真菌组合,复合酶系活性量分别为:低温淀粉酶3000Iu/g~5000Iu/g、蛋白酶1000Iu/g~3000Iu/g、木聚糖酶1000Iu/g~3000Iu/g、果胶酶1000Iu/g~3000Iu/g、纤维素酶1000Iu/g~3000Iu/g。
一种纯固态发酵的低温淀粉酶的生产方法,包括:步骤一,菌种选取;步骤二,菌种培养;步骤三,培养基选取及称量;步骤四,培养基的发酵处理;
其中在上述步骤一中,在产低温淀粉酶为主,包含蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系的单一真菌(如黑曲霉菌或米曲霉菌等)或多种真菌组合作为生产菌株,菌种接种至斜面培养基于20~40℃下培养2~5天后保存备用;
其中在上述步骤二中,将挑选的一种真菌菌株(如黑曲霉菌或米曲霉菌等)或多种真菌菌株组合作为生产菌株接种至培养瓶中,20~40℃下摇瓶培养1~3天,然后移入种子罐中进行通风搅拌培养2~5天;
其中在上述步骤三中,按照复合酶活性量分别为:低温淀粉酶3000Iu/g~5000Iu/g、蛋白酶1000Iu/g~3000Iu/g、木聚糖酶1000Iu/g~3000Iu/g、果胶酶1000Iu/g~3000Iu/g、纤维素酶1000Iu/g~3000Iu/g进行称取;
其中在上述步骤四中,按重量百分比取5~10%的培养菌种接入已消毒的固体发酵培养基中,混匀后加水调整至消毒后含水量为重量百分比50~70%,用乳酸:柠檬酸:富马酸按2:1:1的比重份数进行添加,调节培养基的PH值至4.5~5.5;将培养基逐步放入到底部多孔性托盘内抹平,送入曲房,再自然补给无菌空气,保持环境温度为20~40℃,环境湿度为50~85%的条件下发酵3~7天;每间隔24小时补加一次培养液,其添加量按1%重量比进行均匀喷雾添加;其培养液组成为:糖蜜2%,蛋白胨0.5%,酵母粉0.5%,其余分量为水;发酵成熟的固体料利用低温干燥器进行干燥,直至含水量达到5~12%,再利用粉碎机粉碎至40~60目。
根据上述技术方案,所述步骤四中该培养基的构成为:稻壳粉15~25%,麸皮20~30%,玉米芯8~15%,豆粕10~15%,菜粕4~6%,玉米粉25~35%,糖蜜0.3~0.5%。
根据上述技术方案,所述步骤一中,培养达到生产需要活菌数即单一真菌(如黑曲霉菌或米曲霉菌等)或多种真菌组合≥2±1.5×10的8次方CFU/ml。
根据上述技术方案,所述步骤二中将挑选的一种真菌菌株(如黑曲霉菌或米曲霉菌等)或多种真菌菌株组合接种至培养瓶中,20~40℃下摇瓶培养1~3天,然后移入种子罐中进行通风搅拌培养2~5天;
根据上述技术方案,所述步骤四中将配好的培养基逐步放入到底部多孔性托盘内抹平,送入曲房,再自然补给无菌空气,保持环境温度为20~40℃,环境湿度为50~85%的条件下发酵3~7天。
与现有技术对比,本发明具备以下有益效果:该发明,采用产低温淀粉酶为主、伴生蛋白酶、木聚糖酶、果胶酶和纤维素酶等酶系的单一真菌菌株或多种真菌菌株组合,以稻壳粉、麸皮、玉米、豆粕等作为发酵基料,采用底部多孔性浅盘式固态发酵方法,能够保证菌株快速生长繁殖快,代谢产物量高,其产生的淀粉酶酶系综合活性强,酶系针对性强,全面提升动物饲粮主要能量来源(淀粉)的消化利用率,有利于提升畜禽生产性能和新饲粮的开发利用。该方法也可以缩短培养、发酵周期,提高生产效率,降低生产成本;其还有技术容易掌握、原料来源广泛、对环境无污染、生产规模相对适合等优点。
附图说明
图1为本发明工艺流程图;
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
请参阅图1,本发明提供一种技术方案:一种纯固态发酵的低温淀粉酶的生产方法,配方包括:产低温淀粉酶为主,包含蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系的单一真菌(如黑曲霉菌或米曲霉菌等)或多种真菌组合,复合酶系活性量分别为:低温淀粉酶3000Iu/g~5000Iu/g、蛋白酶1000Iu/g~3000Iu/g、木聚糖酶1000Iu/g~3000Iu/g、果胶酶1000Iu/g~3000Iu/g、纤维素酶1000Iu/g~3000Iu/g。
一种纯固态发酵的低温淀粉酶的生产方法,包括:步骤一,菌种选取;步骤二,菌种培养;步骤三,培养基选取及称量;步骤四,培养基的发酵处理;
其中在上述步骤一中,在产低温淀粉酶为主,伴生蛋白酶、木聚糖酶、果胶酶、纤维素酶等复合酶系的产生菌中挑选出单一真菌菌株(如黑曲霉菌或米曲霉菌等)或多种真菌菌株组合作为生产菌株,以便提高生产效率,降低生产成本,将挑选的菌株接种在斜面上20~40℃下培养2~5天后保存备用;
其中在上述步骤二中,将挑选培养好的斜面菌株分别接入三角培养瓶中在20~40℃的温度条件下摇瓶培养1~3天,再将三角瓶中菌种移入种子罐中进行通风搅拌培养2~5天,培养达到生产需要活菌数即单一真菌(如黑曲霉菌或米曲霉菌等)或多种真菌≥2±1.5×10的8次方CFU/ml;
其中在上述步骤三中,按照复合酶活性量分别为:低温淀粉酶3000Iu/g~5000Iu/g、蛋白酶1000Iu/g~3000Iu/g、木聚糖酶1000Iu/g~3000Iu/g、果胶酶1000Iu/g~3000Iu/g、纤维素酶1000Iu/g~3000Iu/g进行称取;
其中在上述步骤四中,将单一真菌或多种真菌组合按重量百分比取5~10%的培养菌种接入已消毒的固态发酵培养基中,该培养基的构成为:稻壳粉15~25%,麸皮20~30%,玉米芯8~15%,豆粕10~15%,菜粕4~6%,玉米粉25~35%,糖蜜0.3~0.5%,混匀后加水调整至消毒后含水量为重量百分比50~70%,用乳酸:柠檬酸:富马酸按2:1:1的比重份数进行添加,调节培养基的PH值至4.5~5.5;将培养基逐步放入到底部多孔性托盘内抹平,送入曲房,再自然补给无菌空气,保持环境温度为20~40℃,环境湿度为50~85%的条件下发酵3~7天;选用稻壳粉、麸皮、玉米和豆粕等作为发酵基料,有利菌株的生长繁殖和代谢目的产物的获得,可缩短生产周期,在发酵过程中根据菌株生长发酵情况,每间隔24小时补加一次培养液,其添加量按1%重量比进行均匀喷雾添加;其培养液组成为:糖蜜2%,蛋白胨0.5%,酵母粉0.5%,其余分量为水;菌株生长快、长势好的条件下,环境温度、补液和通风调节可控制低一些,反之可控制高一些,视菌株生长代谢情况达到最佳状态,即可停止发酵,发酵成熟的固体料利用低温干燥器进行干燥,直至含水量达到5~12%,再利用粉碎机粉碎至40~60目即可。
从上所述,本发明使用产低温淀粉酶为主,包含蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系的单一真菌菌株(如黑曲霉菌或米曲霉菌等)或多种真菌菌株组合,以稻壳粉、麸皮、玉米、豆粕等作为发酵基料,采用底部多孔性浅盘式纯固态发酵方式,能够保证菌株快速生长繁殖快,酶系形成不受阻遏,制得的低温淀粉酶酶系综合活性与成本性价比高,使用效果显著,技术易掌握、发酵周期短、产品质量稳定,无毒无污染。
实施例1:
下面再以发酵设备试验为实施例对本发明作以详细的说明。
按照复合酶活性量分别为:低温淀粉酶4000Iu/g、蛋白酶2000Iu/g、木聚糖酶2000Iu/g、果胶酶1000Iu/g、纤维素酶1000Iu/g进行称取,先在产低温淀粉酶为主,同时产蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系的产生菌中挑选出一种真菌菌株(如黑曲霉菌或米曲霉菌等)或多种真菌菌株组合,作为生产菌株接种在斜面上,在30℃的温度条件培养48h-62h,然后从斜面菌株接入500ml三角培养瓶中在30℃的温度条件下震荡培养1天,放入装有20L培养液的25L种子罐中进行气流搅拌培养24-48h;全部接入到已消毒的500kg固体发酵培养基中,该培养基的构成为:稻壳粉15%,麸皮25%,豆粕15%,菜粕5%,玉米芯10%,玉米粉30%,糖蜜0.5%,培养基混匀消毒后含水量调整为65%,用乳酸:柠檬酸:富马酸按2:1:1的比重份数进行添加,调节培养基的PH值至4.5-5.5,将500kg固态培养基逐步放入到底部多孔性托盘内抹平,送入曲房,持续补给无菌空气,保持环境温度20~40℃,环境湿度为50~85%之间,间隔24小时补加一次培养液,其添加量按1%重量比进行均匀喷雾添加;其培养液组成为:糖蜜2%,蛋白胨0.5%,酵母粉0.5%,其余分量为水,持续发酵6天,即可停止发酵;发酵成熟的固体料利用低温干燥器进行干燥,直至含水量达到5~12%,再利用粉碎机粉碎至40~60目即可。
实施例2:
选择断奶后健康状况相同的仔猪144头,随机分成两组,即试验组和对照组,每组各6个重复,按实际生产进行饲养管理,试验采用地面平养的方式进行,试验组与对照组每个重复交叉分布;试验组日粮按0.05%比例添加本发明产品,其他与对照组日粮一致,饲养周期38天。
试验期间每天记录投料量,试验开始和结束称量仔猪体重,计算料肉比和成活率。
实验结果如下表:
组别 | 初重(kg) | 末重(kg) | 日增重(g) | 料肉比 | 成活率(%) |
试验组 | 9.54 | 25.26 | 413.68 | 1.51 | 98.6 |
对照组 | 9.78 | 23.89 | 371.32 | 1.64 | 97.2 |
尽管已经出示和描述了本发明的实施例,对于本领域的普通技术人员而言,可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由所附权利要求及其等同物限定。
Claims (6)
1.一种纯固态发酵的低温淀粉酶的生产方法,其特征为:产低温淀粉酶为主,包含蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系,复合酶系活性量分别为:低温淀粉酶3000Iu/g~5000Iu/g、蛋白酶1000Iu/g~3000Iu/g、木聚糖酶1000Iu/g~3000Iu/g、果胶酶1000Iu/g~3000Iu/g、纤维素酶1000Iu/g~3000Iu/g。
2.一种纯固态发酵的低温淀粉酶的生产方法,包括:步骤一,菌种选取;步骤二,菌种培养;步骤三,培养基选取及称量;步骤四,培养基的发酵处理;其特征在于:
其中在上述步骤一中,在产低温淀粉酶为主,包含蛋白酶、木聚糖酶、果胶酶、纤维素酶等伴生酶系的真菌中挑选出一种(如黑曲霉菌或米曲霉菌等)或几种真菌组合作为生产菌株,菌种接种至斜面培养基于20~40℃下培养2~5天后保存备用;
其中在上述步骤二中,将挑选的一种真菌菌株或几种真菌菌株组合接种至培养瓶中,20~40℃下培养1~3天,然后移入种子罐中进行通风搅拌培养2~5天;
其中在上述步骤三中,按照复合酶系活性量分别为:低温淀粉酶3000Iu/g~5000Iu/g、蛋白酶1000Iu/g~3000Iu/g、木聚糖酶1000Iu/g~3000Iu/g、果胶酶1000Iu/g~3000Iu/g、纤维素酶1000Iu/g~3000Iu/g的要求进行培养基称取和混合;
其中在上述步骤四中,按重量百分比取5~10%的培养菌种接入已消毒的固态发酵培养基中,混匀后加水调整至消毒后含水量为重量百分比50~70%,用乳酸:柠檬酸:富马酸按2:1:1的比重份数进行添加,调节培养基的PH值至4.5~5.5;将培养基逐步放入到底部多孔性托盘内抹平,送入曲房,再自然补给无菌空气,保持环境温度为20~40℃,环境湿度为50~85%的条件下发酵3~7天;每间隔24小时补加一次培养液,其添加量按1%重量比进行均匀喷雾添加;其培养液组成为:糖蜜2%,蛋白胨0.5%,酵母粉0.5%,其余分量为水;发酵成熟的固体料利用低温干燥器进行干燥,直至含水量达到5~12%,再利用粉碎机粉碎至40~60目。
3.根据权利要求2所述的一种纯固态发酵的低温淀粉酶的生产方法,其特征在于,所述步骤四中该培养基的构成为:稻壳粉15~25%,麸皮20~30%,玉米芯8~15%,豆粕10~15%,菜粕4~6%,玉米粉25~35%,糖蜜0.3~0.5%。
4.根据权利要求2所述的一种纯固态发酵的低温淀粉酶的生产方法,其特征在于:所述步骤一中,培养达到生产需要活菌数即单一真菌(如黑曲霉菌或米曲霉菌等)或几种真菌组合≥2±1.5×10的8次方CFU/ml。
5.根据权利要求2所述的一种纯固态发酵的低温淀粉酶的生产方法,其特征在于:所述步骤二中将挑选的一种真菌菌株或几种真菌菌株组合接种至培养瓶中,20~40℃下摇瓶培养1~3天,然后移入种子罐中进行通风搅拌培养2~5天。
6.根据权利要求2所述的一种纯固态发酵的低温淀粉酶的生产方法,其特征在于:所述步骤四中将配好的培养基,按要求消毒、接种、调制均匀后逐步放入到底部多孔性托盘内抹平,送入曲房,再自然补给无菌空气,保持环境温度为20~40℃,环境湿度为50~85%的条件下固态发酵3~7天。
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