CN110804600A - 一种浅盘式纯固态发酵复合酶制剂的生产工艺 - Google Patents
一种浅盘式纯固态发酵复合酶制剂的生产工艺 Download PDFInfo
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Abstract
本发明公开了一种浅盘式纯固态发酵复合酶制剂的生产工艺,配方包括:产蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶等复合酶的黑曲霉组合菌群,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,采用高产蛋白酶、木聚糖酶、果胶酶、淀粉酶和纤维素酶等的地衣芽孢杆菌、黑曲霉菌株,稻壳粉、麸皮、玉米芯、菜粕、豆粕等作为发酵基料,能够保证菌株快速生长繁殖快,代谢目的产物的获得,代谢产物量高,活力比现有单一液体发酵技术生产的饲用复合酶提高5倍以上,并且酶组分更全面,各种酶的活力均有不同程度的提高。
Description
技术领域
本发明涉及酶的生产、畜牧养殖技术领域,具体为一种浅盘式纯固态发酵复合酶制剂的生产工艺。
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作用。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。因此设计一种浅盘式纯固态发酵复合酶制剂的生产工艺,充分应用其产生酶系全面,酶系针对性强等特点全面提升外源性复合酶制剂的消化吸收作用,有利于新饲粮的开发利用。
发明内容
本发明的目的在于提供一种浅盘式纯固态发酵复合酶制剂的生产工艺,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:一种浅盘式纯固态发酵复合酶制剂的生产工艺,配方包括:产蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶等复合酶的黑曲霉组合菌群,复合酶活性量分别为:产蛋白酶8000Iu/g、木聚糖酶8000Iu/g、果胶酶2000Iu/g、淀粉酶1000Iu/g、纤维素酶3000Iu/g、甘露聚糖酶500Iu/g、半乳糖苷酶500Iu/g。
一种浅盘式纯固态发酵复合酶制剂的生产工艺,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,在产蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶等产生菌中挑选出地衣芽孢杆菌和黑曲霉作为生产菌株,菌种接种至斜面培养基于20~40℃下培养2~4天后保存备用;
其中在上述步骤二中,将挑选的地衣芽孢杆菌和黑曲霉斜面菌株接种至培养瓶中培养,然后放入种子罐中进行通风搅拌培养1~3天;
其中在上述步骤三中,按照复合酶活性量分别为:产蛋白酶8000Iu/g、木聚糖酶8000Iu/g、果胶酶2000Iu/g、淀粉酶1000Iu/g、纤维素酶3000Iu/g、甘露聚糖酶500Iu/g、半乳糖苷酶500Iu/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长0.5~3米,宽0.5~2米,深5~20cm的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3~5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35~45%,麸皮20~30%,玉米芯8~15%,豆粕8~15%,菜粕4~6%,玉米粉3~5%,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地衣芽孢杆菌≥2±1.5×108次方CFU/ml;黑曲霉菌≥2±1.5×108次方CFU/ml。
根据上述技术方案,所述步骤二中在培养瓶为20~40℃的温度条件下摇瓶培养1~3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长0.5~3米,宽0.5~2米,深5~20cm的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3~5天。
与现有技术对比,本发明具备以下有益效果:该发明,采用高产蛋白酶、木聚糖酶、果胶酶、淀粉酶和纤维素酶等酶系的地衣芽孢杆菌、黑曲霉菌株,稻壳粉、麸皮、豆粕等作为发酵基料,采用底部多孔性浅盘式固态发酵工艺,能够保证菌株快速生长繁殖快,代谢产物量高,活力比现有单一液体发酵技术生产的饲用复合酶提高5倍以上,酶组分更系统全面,各种酶的活力均有不同程度的提高,同时可以缩短培养、发酵周期,提高生产效率,降低生产成本;该方法还具有生产工艺简单、操作简便、技术容易掌握、原料来源广泛、对环境无污染、适合的规模生产等优点。
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:一种浅盘式纯固态发酵复合酶制剂的生产工艺,配方包括:产蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶等复合酶的黑曲霉组合菌群,复合酶活性量分别为:产蛋白酶8000Iu/g、木聚糖酶8000Iu/g、果胶酶2000Iu/g、淀粉酶1000Iu/g、纤维素酶3000Iu/g、甘露聚糖酶500Iu/g、半乳糖苷酶500Iu/g。
一种浅盘式纯固态发酵复合酶制剂的生产工艺,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,在产蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶等复合酶的产生菌中挑选出地衣芽孢杆菌和黑曲霉作为生产菌株,以便提高生产效率,降低生产成本,将挑选的菌株分别接种在斜面上培养成熟;
其中在上述步骤二中,然后将挑选培养好的斜面菌株分别接入三角培养瓶中在20~40℃的温度条件下培养1~3天,再将三角瓶中菌种按重量百分比5%分别放入种子罐中进行通风搅拌培养2~3天,培养达到生产需要活菌数即地衣芽孢杆菌≥2±1.5×108次方CFU/ml;黑曲霉菌≥2±1.5×108次方CFU/ml;
其中在上述步骤三中,按照复合酶活性量分别为:产蛋白酶8000Iu/g、木聚糖酶8000Iu/g、果胶酶2000Iu/g、淀粉酶1000Iu/g、纤维素酶3000Iu/g、甘露聚糖酶500Iu/g、半乳糖苷酶500Iu/g进行称取;
其中在上述步骤四中,将地衣芽胞杆菌:黑曲霉按2:1份量按重量百分比取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35~45%,麸皮20~30%,玉米芯8~15%,豆粕8~15%,菜粕4~6%,玉米粉3~5%,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长0.5~3米,宽0.5~2米,深5~20cm的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3~5天;选用稻壳粉、麸皮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,本发明用高产蛋白酶、木聚糖酶、果胶酶、淀粉酶和纤维素酶的地衣芽孢杆菌、黑曲霉菌株,稻壳粉、麸皮、豆粕等作为发酵基料,采用底部多孔性浅盘式固态发酵工艺,能够保证菌株快速生长繁殖快,酶系形成不受阻遏,制得的饲用复合酶制剂酶活与成本性价比高,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:产蛋白酶8000Iu/g、木聚糖酶8000Iu/g、果胶酶2000Iu/g、淀粉酶1000Iu/g、纤维素酶3000Iu/g、甘露聚糖酶500Iu/g、半乳糖苷酶500Iu/g进行称取,先在产蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶产生菌中挑选出地衣芽孢杆菌、黑曲霉菌株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;全部接入已消毒的500kg固体发酵培养基中,该培养基的构成为:稻壳粉40%,麸皮30%,豆粕10%,菜粕5%,玉米芯10%,玉米粉5%,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长0.5~3米,宽0.5~2米,深5~20cm的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5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。
实施案例2
选择3日龄健康状况相同的三黄鸡2000只,随机分成两组,按实际生产进行饲养管理,试验采用地面平养的方式进行;其中一栋舍为对照组不做任何处理,另一栋为试验组在日粮中按0.03%比例添加本发明产品,饲养50天。
试验期间每天记录投料量,试验开始和结束称鸡体重,计算料肉比和成活率。
实验结果如下表:
尽管已经示出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)
1.一种浅盘式纯固态发酵复合酶制剂,其特征为:含蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶等酶系的纯固态发酵复合酶制剂,复合酶系活性量分别为:产蛋白酶8000Iu/g、木聚糖酶8000Iu/g、果胶酶2000Iu/g、淀粉酶1000Iu/g、纤维素酶3000Iu/g、甘露聚糖酶500Iu/g、半乳糖苷酶500Iu/g。
2.一种浅盘式纯固态发酵复合酶制剂的生产工艺,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,在产含蛋白酶、木聚糖酶、果胶酶、淀粉酶、纤维素酶、半乳糖苷酶、甘露聚糖酶的产生菌中挑选出地衣芽孢杆菌和黑曲霉作为生产菌株,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,将挑选的地衣芽孢杆菌和黑曲霉斜面菌株接种至培养瓶中培养,然后放入种子罐中进行通风搅拌培养1~3天;
其中在上述步骤三中,按照复合酶活性量分别为:产蛋白酶8000Iu/g、木聚糖酶8000Iu/g、果胶酶2000Iu/g、淀粉酶1000Iu/g、纤维素酶3000Iu/g、甘露聚糖酶500Iu/g、半乳糖苷酶500Iu/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长0.5~3米,宽0.5~2米,深5~20cm的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3~5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所述的一种浅盘式纯固态发酵复合酶制剂的生产工艺,其特征在于:所述步骤四中该培养基的构成为:稻壳粉35~45%,麸皮20~30%,玉米芯8~15%,豆粕8~15%,菜粕4~6%,玉米粉3~5%,糖蜜0.3~0.5%。
4.根据权利要求2所述的一种浅盘式纯固态发酵复合酶制剂的生产工艺,其特征在于:所述步骤一中,培养达到生产需要活菌数即地衣芽孢杆菌≥2±1.5×108次方CFU/ml;黑曲霉菌≥2±1.5×108次方CFU/ml。
5.根据权利要求2所述的一种浅盘式纯固态发酵复合酶制剂的生产工艺,其特征在于:所述步骤二中在培养瓶为20~40℃的温度条件下摇瓶培养1~3天。
6.根据权利要求2所述的一种浅盘式纯固态发酵复合酶制剂的生产工艺,其特征在于:所述步骤四中将配好的培养基逐步放入到长0.5~3米,宽0.5~2米,深5~20cm的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3~5天。
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Application publication date: 20200218 |