CN115089672B - 一种饲用发酵中药口服液的制备方法 - Google Patents
一种饲用发酵中药口服液的制备方法 Download PDFInfo
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- CN115089672B CN115089672B CN202210743010.9A CN202210743010A CN115089672B CN 115089672 B CN115089672 B CN 115089672B CN 202210743010 A CN202210743010 A CN 202210743010A CN 115089672 B CN115089672 B CN 115089672B
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- bacillus licheniformis
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Classifications
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- C12R2001/10—Bacillus licheniformis
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
本发明公开了一种饲用发酵中药口服液的制备方法,包括地衣芽孢杆菌活化接种到种子液培养基中发酵,获得地衣芽孢杆菌种子液;地衣芽孢杆菌种子液接种到含有中药粉混合物的液体发酵培养基中发酵,得到地衣芽孢杆菌‑中药粉发酵液混合物;地衣芽孢杆菌‑中药粉发酵液混合物进行离心分离,对上清液进行分离纯化得到饲用发酵中药浓缩液;向饲用发酵中药浓缩液中加入稳定剂进行调配,得到饲用发酵中药口服液。本发明使用特定的地衣芽孢杆菌使得中药粉混合物一同在液体发酵培养基中发酵,利用在发酵过程中的升温和降温替代中药的蒸煮过程,特定的地衣芽孢杆菌在发酵过程中产生的抗菌肽和多种酶系,将中药中的活性物质进一步分解和释放,增强活性。
Description
技术领域
本发明涉及发酵技术领域,特别是涉及一种饲用发酵中药口服液的制备方法。
背景技术
现有技术中,会采用中药提取物来作为饲料添加剂,用于替代促生长类抗生素。但传统中药提取物主要以各种配方中药进行反复煎煮熬制成膏状物或经过后续喷雾干燥制备成粉末,具有成本高,耗能大的缺点,限制了其在畜禽饲料添加剂中的使用。
发明内容
本发明的目的就在于利用地衣芽孢杆菌进行液体发酵能够产生抗菌肽的特点。利用此地衣芽孢杆菌与特定中药粉混合物进行液态深层发酵后,超滤浓缩,调配等工序后,制备得到饲用发酵中药口服液。
为达到上述目的,本发明是按照以下技术方案实施的:
一种饲用发酵中药口服液的制备方法,包括以下步骤:
S1地衣芽孢杆菌种子液制备,将地衣芽孢杆菌活化接种到种子液培养基中发酵,获得地衣芽孢杆菌种子液;
S2地衣芽孢杆菌-中药粉发酵液混合物制备,将步骤S1获得的地衣芽孢杆菌种子液接种到含有中药粉混合物的液体发酵培养基中发酵,得到地衣芽孢杆菌-中药粉发酵液混合物;
S3固液分离,将地衣芽孢杆菌-中药粉发酵液混合物进行离心分离,得到上清液和沉淀;使用超滤膜对上清液进行分离纯化,得到饲用发酵中药浓缩液;
S4饲用发酵中药口服液制备,将步骤S3获得的饲用发酵中药浓缩液加入稳定剂进行调配,得到饲用发酵中药口服液。得到的发酵中药口服液后续会再进行分装,方便使用。
优选的,所述地衣芽孢杆菌(Bacillus licheniformis)保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏时间为2021年12月29日,保藏号为 CGMCC No.24209,保藏单位地址:中国北京。
优选的,所述步骤S1中,地衣芽孢杆菌通过活化扩培制成地衣芽孢杆菌种子液,地衣芽孢杆菌的菌体浓度为1×106-1×109CFU/ml;
每升种子液培养基中包括:蔗糖7-18g、牛骨蛋白胨6-14g、酵母浸粉9-20g、氯化钠0.3-0.9g、硫酸镁0.5-0.9g、硫酸锰0.02-0.09g、柠檬酸三铵3-8g、磷酸氢二钾4-9g,其余为纯水。
进一步的,所述步骤S1中,发酵条件为:pH值为5.0-8.0,发酵温度为 28-42℃,发酵时间为20-44h。
优选的,所述步骤S2中,地衣芽孢杆菌种子液以5-14%的接种量接种至含有中药粉混合物的液体发酵培养基;
每升所述含有中药粉混合物的液体发酵培养基中包括:花生饼粉30-60g、可定性淀粉20-50g、骨蛋白粉5-20g、氯化钠3-8g、磷酸氢二钾4-9g、磷酸二氢钾4-9g、硫酸镁0.8-4g、中药粉混合物80-150g、其余为水;
步骤S2的发酵条件为:pH值为5-8,发酵温度为28-42℃,发酵时间为 20-44h。
进一步的,所述中药粉混合物包括以下重量份的各组分:柴胡5-20份、黄芩10-30份、金银花7-25份、穿心莲15-22份、地棉草8-15份、艾叶10-25份、麦冬5-10份、甘草6-10份、大枣5-10份、麦芽5-15份。
优选的,所述步骤S3中,离心分离时采用碟式离心机,碟式离心机的转速为5000-7000rpm,离心时间15-35min;超滤膜的截留范围为10-50KD。
优选的,所述步骤S4中,稳定剂的加入量为饲用发酵中药口服液质量的 3-10%。
进一步的,所述步骤S4中,稳定剂包括以下重量份的各组分:黄原胶15-30 份、山梨酸钾20-50份、苯甲酸钠15-30份、双乙酸钠15-30份、β-环糊精5-20 份。
进一步的,所述饲用发酵中药口服液的分子量为10-50KD。
本发明的作用原理:
目前,饲料替抗(促生长类抗生素)有两个主要的研究方向,一个是肠道益生菌替抗,另一个是中药提取物替抗,中药提取物中含有大量的有机酸、皂苷、多糖等物质。大量的研究表明地衣芽孢杆菌能够产生抗菌肽,具有改善机体内菌群的组成、提高动物生长性能、增强机体免疫力等综合的保健作用。
依据我国中药炮制学的理论,采用微生物发酵法炮制中药,能够改善中药的口感和提质增效,例如六神曲等。
本发明根据特定的地衣芽孢杆菌(CGMCC No.24209)的特点和中药粉混合物的特性,进行液体深层发酵,制备了饲用发酵中药口服液,为减抗、替抗的路径中进一步提供新的解决方案。
与现有技术相比,本发明的有益效果为:
1)本发明通过使用特定的地衣芽孢杆菌(CGMCC No.24209),使得中药粉混合物一同在液体发酵培养基中发酵,利用液体发酵培养在发酵过程中的升温和降温,替代中药的蒸煮过程,然后利用特定的地衣芽孢杆菌(CGMCC No.24209)在发酵过程中产生的抗菌肽和多种酶系,本发明使用的地衣芽孢杆菌能产生蛋白酶、脂肪酶、淀粉酶和纤维素酶等,将中药中的活性物质进一步分解和释放,增强活性。
2)本发明选用的中药主要是针对气喘和咳嗽等症状,其中柴胡、黄芩、地棉草经过与特定的地衣芽孢杆菌(CGMCC No.24209)发酵后,能够产生协同作用,实现菌药协同的效果,对鸡群的气喘、咳嗽等有着十分明显的调节作用;本发明中的协同作用主要指的是菌酶协同发酵,主要原理是利用本发明的地衣芽孢杆菌(菌)和其代谢产物产生的多种酶系(纤维素酶、蛋白酶、脂肪酶、淀粉酶等)进行组合发酵,此发酵方式属于菌酶协同耗氧发酵。
3)采用液体发酵的形式可以连续生产,机械化程度高,操作重复性好,与传统的中药添加剂相比,使用更方便,作用更迅速,生物利用度更高。
附图说明
图1为本发明的工艺流程图。
具体实施方式
下面以具体实施例对本发明作进一步描述,在此发明的示意性实施例以及说明用来解释本发明,但并不作为对本发明的限定。
下述实施例中使用的设备型号如下:发酵罐为江苏丰泽生物工程设备制造有限公司生产的型号为FN-E型100L;碟式离心机为江苏巨能机械有限公司生产的型号:DBY211。
实施例1
如图1所示,一种饲用发酵中药口服液的制备方法,包括以下步骤,
S1地衣芽孢杆菌种子液制备,将地衣芽孢杆菌活化接种到种子液培养基中发酵,获得地衣芽孢杆菌种子液;
地衣芽孢杆菌(Bacillus licheniformis)保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏时间为2021年12月29日,保藏号为CGMCC No.24209,保藏单位地址:中国北京;
地衣芽孢杆菌的菌体浓度为1.2×106CFU/ml,发酵条件为:pH值为6.2,发酵温度为32℃,发酵时间为35h;
每升种子液培养基中包括:蔗糖7g、牛骨蛋白胨6g、酵母浸粉9g、氯化钠 0.3g、硫酸镁0.5g、硫酸锰0.02g、柠檬酸三铵3g、磷酸氢二钾4g,其余为纯水。
S2地衣芽孢杆菌-中药粉发酵液混合物制备,将步骤S1获得的地衣芽孢杆菌种子液以5%的接种量,接种到含有中药粉混合物的液体发酵培养基中发酵,得到地衣芽孢杆菌-中药粉发酵液混合物;
每升含有中药粉混合物的液体发酵培养基中包括:花生饼粉30g、可定性淀粉20g、骨蛋白粉5g、氯化钠3g、磷酸氢二钾4g、磷酸二氢钾4g、硫酸镁0.8g、中药粉混合物80g、其余为水。中药粉混合物包括以下重量份的各组分:柴胡5 份、黄芩25份、金银花9份、穿心莲22份、地棉草8份、艾叶10份、麦冬5 份、甘草6份、大枣5份、麦芽5份。
发酵条件为:pH值为6.3,发酵温度为40℃,发酵时间为28h。
S3固液分离,将地衣芽孢杆菌-中药粉发酵液混合物在碟式离心机中离心分离,得到上清液和沉淀;使用超滤膜对上清液进行分离纯化,得到饲用发酵中药浓缩液;
碟式离心机的转速为5000rpm,离心时间15min。超滤膜的截留范围为 30KD。
S4饲用发酵中药口服液制备,将步骤S3获得的饲用发酵中药浓缩液,得到饲用发酵中药口服液;稳定剂的加入量为饲用发酵中药口服液质量的3%,得到的发酵中药口服液后续会再进行分装,方便使用;
稳定剂包括以下重量份的各组分:黄原胶20份、山梨酸钾30份、苯甲酸钠15份、双乙酸钠30份、β-环糊精5份。
本实施例获得的饲用发酵中药口服液的分子量为30KD,易被机体吸收利用。
实施例2
一种饲用发酵中药口服液的制备方法,包括以下步骤,
S1地衣芽孢杆菌种子液制备,将地衣芽孢杆菌活化接种到种子液培养基中发酵,获得地衣芽孢杆菌种子液;
地衣芽孢杆菌(Bacillus licheniformis)保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏时间为2021年12月29日,保藏号为CGMCC No.24209,保藏单位地址:中国北京;
地衣芽孢杆菌的菌体浓度为1.6×108CFU/ml;发酵条件为:pH值为7.5,发酵温度为39℃,发酵时间为40h。
每升种子液培养基中包括:蔗糖18g、牛骨蛋白胨14g、酵母浸粉20g、氯化钠0.9g、硫酸镁0.9g、硫酸锰0.09g、柠檬酸三铵8g、磷酸氢二钾9g,其余为纯水。
S2地衣芽孢杆菌-中药粉发酵液混合物制备,将步骤S1获得的地衣芽孢杆菌种子液以14%的接种量,接种到含有中药粉混合物的液体发酵培养基中发酵,得到地衣芽孢杆菌-中药粉发酵液混合物;
每升含有中药粉混合物的液体发酵培养基中包括:花生饼粉45g、可定性淀粉30g、骨蛋白粉15g、氯化钠8g、磷酸氢二钾9g、磷酸二氢钾9g、硫酸镁 4g、中药混合物150g、其余为水。中药粉混合物包括以下重量份的各组分:柴胡15份、黄芩10份、金银花10份、穿心莲19份、地棉草8份、艾叶12份、麦冬7份、甘草7份、大枣6份、麦芽6份。
发酵条件为:pH值为7.6,发酵温度为42℃,发酵时间为36h,
S3固液分离,将地衣芽孢杆菌-中药粉发酵液混合物在碟式离心机中离心,得到上清液和沉淀;使用超滤膜对上清液进行分离纯化,得到饲用发酵中药浓缩液;碟式离心机的转速为6500rpm,离心时间25min;超滤膜的截留范围为 50KD。
S4饲用发酵中药口服液制备,将步骤S3获得的饲用发酵中药浓缩液,得到饲用发酵中药口服液;稳定剂的加入量为饲用发酵中药口服液质量的6%,得到的发酵中药口服液后续会再进行分装,方便使用;
稳定剂包括以下重量份的各组分:黄原胶25份、山梨酸钾25份、苯甲酸钠20份、双乙酸钠20份、β-环糊精10份。
本实施例获得的饲用发酵中药口服液的分子量为50KD,易被机体吸收利用。
试验
本发明实施例1获得的饲用发酵中药口服液的使用方法:
每600kg饮水加饲用发酵中药口服液500ml,集中投喂。每个疗程7-12 天,每个疗程间隔2-4天,可进入下一个疗程。正常情况下,2个疗程能明显见效。然后再使用一个疗程,巩固疗效。
具体案例:
(1)2021年10月洛阳市宜阳县赵保镇王海养鸡场,200日龄,该鸡场养殖规模是10000只。主要存在的问题是出现有轻微的气喘和咳嗽等症状。该鸡场使用实施例1获得的饲用发酵中药口服液第5天,气喘和咳嗽症状恢复正常的水平达到80-90%,鸡群的状态基本恢复到正常的状态。使用15天后,基本恢复正常水平。
(2)2021年12月新乡市辉县王志忠养鸡场,210日龄,该鸡场养殖规模是15000只。主要存在的问题是气喘和咳嗽,粪便稀,有腹泻。该鸡场使用饲用实施例1获得的发酵中药口服液第6天,气喘和咳嗽症状恢复正常的水平达到85-95%,粪便稀和腹泻等症状基本消失。继续使用7天后,鸡群基本恢复正常水平。
由此可知,本发明或的饲用发酵中药口服液能够对鸡气喘和咳嗽症状有很好的预防功能(本发明获得的饲用发酵中药口服液预防为主,治疗为辅,主要在家禽发生症状的前期使用,治疗前期病症,并预防病症的进一步发展),减少鸡在养殖过程中抗生素的使用量。
本发明的技术方案不限于上述具体实施例的限制,凡是根据本发明的技术方案做出的技术变形,均落入本发明的保护范围之内。
序列表
<110> 焦作市佰役安生物工程有限公司
<120> 一种饲用发酵中药口服液的制备方法
<141>
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1488
<212> DNA
<213> 地衣芽孢杆菌CGMCC No.24209(Bacillus licheniformis)
<400> 1
ggctcaggac gaacgctggc ggcgtgccta atacatgcaa gtcgagcgga cagatgggag 60
cttgctccct gatgttagcg gcggacgggt gagtaacacg tgggtaacct gcctgtaaga 120
ctgggataac tccgggaaac cggggctaat accggatgct tgattgaacc gcatggttca 180
attataaaag gtggctttta gctaccactt acagatggac ccgcggcgca ttagctagtt 240
ggtgaggtaa cggctcacca aggcaacgat gcgtagccga cctgagaggg tgatcggcca 300
cactgggact gagacacggc ccagactcct acgggaggca gcagtaggga atcttccgca 360
atggacgaaa gtctgacgga gcaacgccgc gtgagtgatg aaggttttcg gatcgtaaaa 420
ctctgttgtt agggaagaac aagtaccgtt cgaatagggc ggtaccttga cggtacctaa 480
ccagaaagcc acggctaact acgtgccagc agccgcggta atacgtaggt ggcaagcgtt 540
gtccggaatt attgggcgta aagcgcgcgc aggcggtttc ttaagtctga tgtgaaagcc 600
cccggctcaa ccggggaggg tcattggaaa ctggggaact tgagtgcaga agaggagagt 660
ggaattccac gtgtagcggt gaaatgcgta gagatgtgga ggaacaccag tggcgaaggc 720
gactctctgg tctgtaactg acgctgaggc gcgaaagcgt ggggagcgaa caggattaga 780
taccctggta gtccacgccg taaacgatga gtgctaagtg ttagagggtt tccgcccttt 840
agtgctgcag caaacgcatt aagcactccg cctggggagt acggtcgcaa gactgaaact 900
caaaggaatt gacgggggcc cgcacaagcg gtggagcatg tggtttaatt cgaagcaacg 960
cgaagaacct taccaggtct tgacatcctc tgacaaccct agagataggg cttccccttc 1020
gggggcagag tgacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt 1080
aagtcccgca acgagcgcaa cccttgatct tagttgccag cattcagttg ggcactctaa 1140
ggtgactgcc ggtgacaaac cggaggaagg tggggatgac gtcaaatcat catgcccctt 1200
atgacctggg ctacacacgt gctacaatgg gcagaacaaa gggcagcgaa gccgcgaggc 1260
taagccaatc ccacaaatct gttctcagtt cggatcgcag tctgcaactc gactgcgtga 1320
agctggaatc gctagtaatc gcggatcagc atgccgcggt gaatacgttc ccgggccttg 1380
tacacaccgc ccgtcacacc acgagagttt gtaacacccg aagtcggtga ggtaaccttt 1440
tggagccagc cgccgaaggt gggacagatg attggggtga agtcgtaa 1488
Claims (6)
1.一种饲用发酵中药口服液的制备方法,其特征在于,包括以下步骤:
S1地衣芽孢杆菌种子液制备,将地衣芽孢杆菌活化接种到种子液培养基中发酵,获得地衣芽孢杆菌种子液;
S2地衣芽孢杆菌-中药粉发酵液混合物制备,将步骤S1获得的地衣芽孢杆菌种子液接种到含有中药粉混合物的液体发酵培养基中发酵,得到地衣芽孢杆菌-中药粉发酵液混合物;
S3固液分离,将地衣芽孢杆菌-中药粉发酵液混合物进行离心分离,得到上清液和沉淀;使用超滤膜对上清液进行分离纯化,得到饲用发酵中药浓缩液;
S4饲用发酵中药口服液制备,将步骤S3获得的饲用发酵中药浓缩液加入稳定剂进行调配,得到饲用发酵中药口服液;
所述地衣芽孢杆菌保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏时间为2021年12月29日,保藏号为CGMCC No.24209;
所述步骤S2中,地衣芽孢杆菌种子液以5-14%的接种量接种至含有中药粉混合物的液体发酵培养基;
每升所述含有中药粉混合物的液体发酵培养基中包括:花生饼粉30-60g、可溶性淀粉20-50g、骨蛋白粉5-20g、氯化钠 3-8g、磷酸氢二钾 4-9g、磷酸二氢钾 4-9g、硫酸镁0.8-4g、中药粉混合物80-150g、其余为水;
步骤S2的发酵条件为:pH值为5-8,发酵温度为28-42℃,发酵时间为20-44h;
所述中药粉混合物由以下重量份的各组分制成:柴胡5-20份、黄芩10-30份 、金银花7-25份、穿心莲15-22份、地棉草8-15份、艾叶10-25份、麦冬5-10份、甘草6-10份、大枣5-10份、麦芽5-15份。
2.根据权利要求1所述的一种饲用发酵中药口服液的制备方法,其特征在于:所述步骤S1中,地衣芽孢杆菌的菌体浓度为1×106-1×109CFU/ml;
每升种子液培养基中包括:蔗糖7-18g、牛骨蛋白胨6-14g、酵母浸粉9-20g、氯化钠0.3-0.9g、硫酸镁0.5-0.9g、硫酸锰0.02-0.09g、柠檬酸三铵3-8g、磷酸氢二钾4-9g,其余为纯水。
3.根据权利要求2所述的一种饲用发酵中药口服液的制备方法,其特征在于:所述步骤S1中,发酵条件为:pH值为5.0-8.0,发酵温度为28-42℃,发酵时间为20-44h。
4.根据权利要求1所述的一种饲用发酵中药口服液的制备方法,其特征在于:所述步骤S3中,离心分离时采用碟式离心机,碟式离心机的转速为5000-7000rpm,离心时间15-35min;超滤膜的截留范围为10-50KD。
5.根据权利要求1所述的一种饲用发酵中药口服液的制备方法,其特征在于:所述步骤S4中,稳定剂的加入量为饲用发酵中药口服液质量的3-10%。
6.根据权利要求5所述的一种饲用发酵中药口服液的制备方法,其特征在于:所述步骤S4中,稳定剂包括以下重量份的各组分:黄原胶15-30份、山梨酸钾20-50份、苯甲酸钠15-30份、双乙酸钠15-30份、β-环糊精5-20份。
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Denomination of invention: A method for preparing fermented traditional Chinese medicine oral liquid for feeding Granted publication date: 20231222 Pledgee: Bank of China Limited by Share Ltd. Jiaozuo branch Pledgor: Jiaozuo Baiyian Bioengineering Co.,Ltd. Registration number: Y2024980009406 |