CN115606678A - 一种复合饲用益生菌利用废弃羽毛制备高蛋白益生动物饲料的方法 - Google Patents
一种复合饲用益生菌利用废弃羽毛制备高蛋白益生动物饲料的方法 Download PDFInfo
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Abstract
本发明公开了一种复合饲用益生菌利用废弃羽毛制备高蛋白益生动物饲料的方法。其过程是将复合菌剂种子液添加至含有羽毛的发酵培养基,经高密度发酵培养后获得混合液;以混合液的体积比(m/v)向混合液中添加甘蔗渣5~10%、海藻粉5~10%、麦麸8~12%、淀粉7~10%,混匀后造粒,获得高蛋白益生动物饲料;所述的复合菌剂为含有枯草芽孢杆菌、地衣芽孢杆菌和片球菌的组合。本发明的高蛋白益生动物饲料具有益生、替抗和改善适口性功能,利于提升饲料的利用率,同时具有缓解我国蛋白饲料资源供需矛盾的潜力,助力我国养殖业与饲料领域的持续健康发展。
Description
技术领域:
本发明属于饲料技术领域,具体涉及了一种复合益生菌利用废弃羽毛制备高蛋白益生动物饲料的方法。
背景技术:
羽毛是畜禽养殖业的主要副产物之一,约占鸟类和禽类体重的10%左右。羽毛因富含角蛋白(在85%以上)被视为一种天然的高蛋白资源。养殖业的集约化快速发展使我国每年产生数百万吨的废弃羽毛。目前,研究者对大部分羽毛只进行了简单处理(如填埋、焚烧、酶解)导致羽毛未能被充分利用,这无疑造成了蛋白资源(废弃羽毛)浪费和环境污染。因我国蛋白饲料资源短缺而长期依赖进口,造成蛋白饲料资源的供求关系紧张,限制了我国养殖业与饲料领域的规模化发展。而高含蛋白的废弃羽毛用作蛋白饲料将有效缓解我国蛋白饲料资源的供需矛盾。因此,废弃羽毛的饲料化应用受到了广泛关注。
由于羽毛结构中存在氢键、疏水作用和丰富的二硫键等作用力,导致其结构致密而难以降解,这也是羽毛未能被充分利用的限制因素。大量的研究发现普通的酶(如胰蛋白酶、胃蛋白酶)很难降解羽毛。微生物产生的角蛋白酶具有降解羽毛的能力,且以枯草芽孢杆菌和地衣芽孢杆菌的报道最多。因单菌株产角蛋白酶能力有限而不能高效降解羽毛,而复合菌能产生种类丰富的蛋白酶,有利于打开羽毛结构中的多种共价键,特别是二硫键。研究发现,羽毛是在以角蛋白酶为主的多种酶系作用下实现降解的。相比单菌株,多株菌构建的复合菌将增强降解羽毛的能力。此外,饲用益生菌能产生抗动物肠道病菌的活性物质,且具有益生功能,减少动物疾病,目前已用作饲料添加剂。而采用复合益生菌直接且高效降解废弃羽毛尚未见报道。因此,复合饲用益生菌对加快废弃羽毛的降解及其饲料化与资源化应用具有重要的经济价值。
发明内容
本发明的目的在于提供一种复合饲用益生菌利用废弃羽毛制备高蛋白益生动物饲料的方法。本发明以枯草芽孢杆菌、地衣芽孢杆菌、片球菌构建复合菌进行降解废弃羽毛并制备高蛋白益生动物饲料,且饲料中可溶性蛋白含量在36.3g/L左右、益生菌活菌数在2.85×108cfu/g左右。
本发明的复合饲用益生菌利用废弃羽毛制备高蛋白益生动物饲料的方法,包括以下步骤:
将复合菌剂种子液添加至含有羽毛的发酵培养基,经高密度发酵培养后获得羽毛降解物、复合益生菌体及其代谢物的混合液,向上述混合液中添加甘蔗渣10%、海藻粉8%、麦麸10%和淀粉10%(均为混合液的体积比,m/v,g/ml),混匀后造粒,获得高蛋白益生动物饲料,其直径约为4mm,长度为6~12mm。所述的复合菌剂种子液为含有枯草芽孢杆菌、地衣芽孢杆菌和片球菌的组合。
所述的枯草芽孢杆菌、地衣芽孢杆菌和片球菌都是从LB培养基平板活化菌株,分别接种至液体LB培养基培养12~36h制备种子液,然后将枯草芽孢杆菌、地衣芽孢杆菌和片球菌种子液按照体积比1-4:1-3:1复配得到复合菌剂种子液。
获得的种子液和复合菌剂种子液,分别以5~12%接种量转接至筛选培养基中,在150~250r/min和37℃条件下培养20~48h,通过差重法比较各菌株降解废弃羽毛的效率。
所述的筛选培养基,按质量分数计包括:0.06~0.09%NaCl、0.06~0.1%KH2PO4、0.01~0.09%K2HPO4、0.002~0.005%MgSO4·7H2O、1%羽毛或1%羽毛粉,pH为7.0~10,121℃灭菌20min。
进一步优选,是按照体积比2:2:1将枯草芽孢杆菌、地衣芽孢杆菌和片球菌种子液复配得到复合菌剂种子液。
所述的羽毛优选为膨化羽毛粉,是先用双螺杆挤压膨化机处理废弃羽毛,获得粗羽毛粉;其中,膨化机的转速为160~180r/min、温度为158~164℃、压力为0.38~0.45MPa;再经200目过滤筛过滤,获得精细均一的羽毛粉。
优选,是将复合菌剂种子液按照体积比4~18%的接种量转接至含有膨化羽毛粉的发酵培养基中,在30~50℃和150~250r/min条件下降解羽毛粉;经补加葡萄糖和膨化羽毛粉进行高密度发酵2~5天,获得羽毛降解物、复合益生菌及其代谢物的混合液;所述的发酵培养基,按质量分数计包括:0.01~0.09%NaCl、0.01~0.1%KH2PO4、0.02~0.1%K2HPO4、0.002~0.01%MgSO4·7H2O,1~9%羽毛粉,pH为7.0~10,121℃灭菌20min。
优选,所述的造粒是在235~245r/min条件下进行造粒,获得饲料,饲料的直径约为4mm,长度为6~12mm。
优选,采用凯氏定氮法(GB/T 6432-2018)测定饲料中的粗蛋白和可溶性蛋白含量,过滤法(GB/T 6434-2006)测定粗纤维含量,分光光度法检测角蛋白酶活力(全饲标[2021]52号)。
现有技术相比,本发明具有以下优点:
1、与单菌、两菌株联合作用相比,复合益生菌处理羽毛量大,产生可溶性蛋白的含量高,易被禽畜动物吸收,提高饲料利用率。
2、饲料中活性益生菌数量在2.85×108cfu/g以上。
3、高蛋白益生动物饲料中复合益生菌的益生功能更全面,有利于提高免疫力,减少疾病发生,推进抗生素的替代进程。
4、三株菌的复合显著提高角蛋白酶活力。
Bacillus subtilis HC2,其于2022年5月12日保藏于广东省微生物菌种保藏中心(GDMCC),地址:广州市先烈中路100号大院59号楼5楼,邮编:510070,保藏号GDMCC No:62466。
附图说明
图1是菌种复配比对羽毛粉降解的影响;
图2是可溶性蛋白与角蛋白酶活力的比较,a:复合菌发酵前后的可溶性蛋白含量,b:单菌株与复合菌发酵液中的角蛋白酶活力;
图3是复合益生菌发酵前后的菌种占比的变化;
图4是复合益生菌高蛋白动物饲料。
具体实施方式
以下实施例是对本发明的进一步说明,而不是对本发明的限制。
枯草芽孢杆菌可以是常规的枯草芽孢杆菌,本实施例选择Bacillus subtilisHC2,其于2022年5月12日保藏于广东省微生物菌种保藏中心(GDMCC),地址:广州市先烈中路100号大院59号楼5楼,邮编:510070,保藏号GDMCC No:62466。
地衣芽孢杆菌可以是常规的地衣芽孢杆菌,本实施例选择地衣芽孢杆菌Lu2。
片球菌可以是常规戊糖片球菌,本实施例选择戊糖片球菌PU4。
以下实施例使用的羽毛粉的制备方法是鸡毛经双螺杆挤压膨化机处理(含水量为15~18%),获得粗羽毛粉,膨化机的转速为160~180r/min、温度为158~164℃、压力为0.38~0.45MPa;再经200目过滤筛过滤,获得精细均一的羽毛粉,备用。
实施例1三株菌对经预处理废弃羽毛降解效果的比较
在固态LB培养基中活化枯草芽孢杆菌HC2(保藏号为GDMCC No.62466)、地衣芽孢杆菌Lu2与戊糖片球菌PU4三株菌,分别接种至液体LB培养基培养16h,制备种子液;以体积比6%接种量接种至筛选培养基,在200r/min和37℃条件下培养30h,采用差重法检测其降解羽毛(粉)的效率。如表1所示,三株菌对经高温灭菌处理废弃羽毛的降解率均在34%以下,降解羽毛能力大小顺序为Lu2菌>HC2菌>PU4菌;而它们对经膨化-高温灭菌处理废弃羽毛(121℃,20min)的降解效率显著升高,均在80%左右。表明膨化处理能够加快废弃羽毛的降解。
液态LB培养基:按质量分数计,包括0.5%酵母粉、1%胰蛋白胨、1%氯化钠(固态时则需添加1.5%琼脂),pH 7.0,溶剂为水,其配置方法是将各成分混合均匀,经121℃灭菌20min。
筛选培养基:按质量分数计,包括0.07%NaCl、0.09%KH2PO4、0.03%K2HPO4、0.004%MgSO4·7H2O、1%羽毛或1%羽毛粉,pH为8.5,溶剂为水,其配置方法是将各成分混合均匀,经121℃灭菌20min。
表1三株菌对预处理废弃羽毛的降解效果比较
实施例2复合益生菌高密度发酵制备高蛋白益生动物饲料
将实施例1的Lu2菌、HC2菌和PU4菌的种子液以体积比(1~4):(1~3):1混匀,以体积比6%的总接种量转接至含1%羽毛粉的筛选培养基,在200r/min和37℃条件下培养30h,比较不同复配比的复合益生菌对羽毛粉的降解效率。如图1所示,复配比为2:2:1时复合菌降解羽毛粉的效率达到最高(>92%)。按复配比为体积比2:2:1将Lu2菌、HC2菌和PU4菌的种子液复配,获得复合菌剂种子液。以培养基体积的8%为总接种量将其转接至含6%羽毛粉的发酵培养基(500mL/L),在40℃、200r/min和pH 8.5条件下降解羽毛粉,经分批补加发酵液的0.5%(m/v)葡萄糖和6%(m/v)羽毛粉进行复合菌的高密度发酵4天,获得羽毛降解物、复合益生菌体及其功能代谢产物的混合液;以单株菌的种子液按照相同处理作为对照。该混合液中可溶性蛋白含量达到36.3g/L,较发酵前(7.55g/L)提高了4.8倍(图2a);角蛋白酶活力达到1857U/mL,是单菌株产角蛋白酶活力的3~5.7倍(图2b)。与发酵前菌落数相比,混合液中复合益生菌活菌数明显增加,达到了4.33×108cfu/mL;Lu2菌、HC2菌和PU4菌活菌数均有显著增加,同时复合菌的菌株组成也发生了变化:Lu2菌和PU4菌在复合菌中所占比例增大,而HC2菌所占比例则下降(图3)。这可能是复合菌中菌株间的互作关系引起的变化,其中PU4菌产生的乳酸对羽毛/羽毛粉具有一定的软化作用。这表明复合菌能以膨化羽毛作为唯一的碳源和氮源进行生长代谢,且与单菌相比产生更高活性的角蛋白酶,加速羽毛(粉)降解,从而获得高浓度的可溶性蛋白和活性益生菌。
以混合液的质量体积比向上述混合液中添加甘蔗渣10%(m/v,g/ml)、海藻粉8%(m/v,g/ml)、麦麸10%(m/v,g/ml)和淀粉10%(m/v,g/ml),混匀后造粒,制得饲料。如图4所示,饲料直径约为4mm,长度为6~12mm;经检测发现饲料中粗蛋白含量为37%、粗纤维含量在8.2%左右、益生菌活菌数达到3.2×108cfu/g。该复合益生菌饲料保持较高的益生菌活菌数,高于Q/HBMSY 001-2019标准的复合益生菌发酵饲料中益生菌的数量,符合高蛋白益生动物饲料的特征。
筛选培养基与实施例1中相同,其中添加的是1%羽毛粉。
发酵培养基:按质量分数计,包括0.06%NaCl、0.05%KH2PO4、0.03%K2HPO4、0.004%MgSO4·7H2O,6%羽毛粉,pH 8.5,121℃灭菌20min。
Claims (8)
1.一种复合饲用益生菌利用废弃羽毛制备高蛋白益生动物饲料的方法,其特征在于,包括以下步骤:
将复合菌剂种子液添加至含有羽毛的发酵培养基,经发酵培养后获得羽毛降解物、复合益生菌体及其代谢物的混合液;以混合液的质量体积比(m/v,g/ml)向混合液中添加甘蔗渣5~10%、海藻粉5~10%、麦麸8~12%、淀粉7~10%,混匀后造粒,获得高蛋白益生动物饲料,所述的复合菌剂种子液为含有枯草芽孢杆菌、地衣芽孢杆菌和片球菌的组合。
2.根据权利要求1所述的方法,其特征在于,所述的枯草芽孢杆菌、地衣芽孢杆菌和片球菌都是从LB培养基平板活化菌株,分别接种至液体LB培养基培养12~36h制备种子液,然后将枯草芽孢杆菌、地衣芽孢杆菌和片球菌种子液按照体积比1-4:1-3:1的比例复配而得到复合菌剂种子液。
3.根据权利要求2所述的方法,其特征在于,所述的种子液和复合菌剂种子液,均以5~12%接种量转接至筛选培养基中,在150~250r/min和37℃条件下培养20~48h,所述的筛选培养基,按质量分数计包括:0.06~0.09%NaCl、0.06~0.1%KH2PO4、0.01~0.09%K2HPO4、0.002~0.005%MgSO4·7H2O、1%羽毛或1%羽毛粉,pH为7.0~10,121℃灭菌20min。
4.根据权利要求2、3所述的方法,其特征在于,是按照体积比2:2:1将枯草芽孢杆菌、地衣芽孢杆菌和片球菌种子液复配得到复合菌剂种子液。
5.根据权利要求1所述的方法,其特征在于,所述的羽毛为膨化羽毛粉,是先用双螺杆挤压膨化机处理废弃羽毛,获得粗羽毛粉;其中,膨化机的转速为160~180r/min、温度为158~164℃、压力为0.38~0.45MPa;再经200目过滤筛过滤,获得精细均一的羽毛粉。
6.根据权利要求1、4所述的方法,其特征在于,是将复合菌剂种子液按照体积比4~18%的接种量转接至含有膨化羽毛粉的发酵培养基中,在30~50℃和pH 7~11条件下降解羽毛粉;经补加葡萄糖和膨化羽毛粉进行发酵2~5天,获得混合液;所述的发酵培养基,按质量分数计包括:0.01~0.09%NaCl、0.01~0.1%KH2PO4、0.02~0.1%K2HPO4、0.002~0.01%MgSO4·7H2O,1~9%羽毛粉,pH为7.0~10,121℃灭菌20min。
7.根据权利要求1所述的方法,其特征在于,所述的废弃羽毛为鸡毛、鸭毛、鹅毛。
8.根据权利要求1所述的方法,是采用复合益生菌将废弃羽毛转化为高蛋白益生动物饲料。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102389022A (zh) * | 2011-08-11 | 2012-03-28 | 湖北大学 | 利用地衣芽孢杆菌降解羽毛制备菌肽复合蛋白饲料添加剂的方法 |
CN102726596A (zh) * | 2012-06-18 | 2012-10-17 | 杭州启明星生物营养有限公司 | 一种羽毛蛋白肽饲料添加剂及应用 |
CN105614023A (zh) * | 2015-12-31 | 2016-06-01 | 湖州环农微生物技术研究所 | 一种用于豆粕发酵的发酵酶解剂及其应用 |
CN106173213A (zh) * | 2016-07-06 | 2016-12-07 | 中国科学院合肥物质科学研究院 | 一种鱼粉的制作方法 |
CN115067435A (zh) * | 2022-06-24 | 2022-09-20 | 广东省科学院生物与医学工程研究所 | 一种以废弃羽毛、果皮及裂殖壶菌菌渣为原料的高蛋白鸽子饲料及其制备方法 |
-
2022
- 2022-10-08 CN CN202211222993.8A patent/CN115606678A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102389022A (zh) * | 2011-08-11 | 2012-03-28 | 湖北大学 | 利用地衣芽孢杆菌降解羽毛制备菌肽复合蛋白饲料添加剂的方法 |
CN102726596A (zh) * | 2012-06-18 | 2012-10-17 | 杭州启明星生物营养有限公司 | 一种羽毛蛋白肽饲料添加剂及应用 |
CN105614023A (zh) * | 2015-12-31 | 2016-06-01 | 湖州环农微生物技术研究所 | 一种用于豆粕发酵的发酵酶解剂及其应用 |
CN106173213A (zh) * | 2016-07-06 | 2016-12-07 | 中国科学院合肥物质科学研究院 | 一种鱼粉的制作方法 |
CN115067435A (zh) * | 2022-06-24 | 2022-09-20 | 广东省科学院生物与医学工程研究所 | 一种以废弃羽毛、果皮及裂殖壶菌菌渣为原料的高蛋白鸽子饲料及其制备方法 |
Non-Patent Citations (1)
Title |
---|
江永才: "发酵饲料生产现状浅析", 畜禽业, no. 03, pages 34 - 36 * |
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