CN103343101A - 土霉素菌种斜面培养基 - Google Patents
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- KIPLYOUQVMMOHB-MXWBXKMOSA-L [Ca++].CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O.CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O Chemical compound [Ca++].CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O.CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O KIPLYOUQVMMOHB-MXWBXKMOSA-L 0.000 title claims abstract description 29
- 229940063650 terramycin Drugs 0.000 title claims abstract description 29
- 239000001963 growth medium Substances 0.000 title abstract description 7
- 239000000843 powder Substances 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229920001817 Agar Polymers 0.000 claims abstract description 12
- 239000001888 Peptone Substances 0.000 claims abstract description 12
- 108010080698 Peptones Proteins 0.000 claims abstract description 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 12
- 229920002472 Starch Polymers 0.000 claims abstract description 12
- 239000008272 agar Substances 0.000 claims abstract description 12
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims abstract description 12
- 235000019319 peptone Nutrition 0.000 claims abstract description 12
- 235000019698 starch Nutrition 0.000 claims abstract description 12
- 239000008107 starch Substances 0.000 claims abstract description 12
- 239000002994 raw material Substances 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims description 14
- 229910019142 PO4 Inorganic materials 0.000 claims description 11
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 11
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 11
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 11
- 239000010452 phosphate Substances 0.000 claims description 11
- 229910052700 potassium Inorganic materials 0.000 claims description 11
- 239000011591 potassium Substances 0.000 claims description 11
- 230000001580 bacterial effect Effects 0.000 abstract description 17
- 241000209140 Triticum Species 0.000 abstract description 6
- 235000021307 Triticum Nutrition 0.000 abstract description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 abstract 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 abstract 2
- 238000009395 breeding Methods 0.000 abstract 1
- 230000001488 breeding effect Effects 0.000 abstract 1
- 230000007547 defect Effects 0.000 abstract 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 abstract 1
- 235000019797 dipotassium phosphate Nutrition 0.000 abstract 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 abstract 1
- 235000019341 magnesium sulphate Nutrition 0.000 abstract 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 abstract 1
- 235000019796 monopotassium phosphate Nutrition 0.000 abstract 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 239000011780 sodium chloride Substances 0.000 abstract 1
- 235000015099 wheat brans Nutrition 0.000 description 12
- 239000002609 medium Substances 0.000 description 5
- 244000061458 Solanum melongena Species 0.000 description 4
- 235000002597 Solanum melongena Nutrition 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
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- 238000000855 fermentation Methods 0.000 description 3
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- 241000894006 Bacteria Species 0.000 description 2
- 241000187419 Streptomyces rimosus Species 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 239000004100 Oxytetracycline Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 229960000625 oxytetracycline Drugs 0.000 description 1
- IWVCMVBTMGNXQD-PXOLEDIWSA-N oxytetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3[C@H](O)[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-PXOLEDIWSA-N 0.000 description 1
- 235000019366 oxytetracycline Nutrition 0.000 description 1
- 230000035040 seed growth Effects 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- IWVCMVBTMGNXQD-UHFFFAOYSA-N terramycin dehydrate Natural products C1=CC=C2C(O)(C)C3C(O)C4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-UHFFFAOYSA-N 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种土霉素菌种斜面培养基，其特征在于：它由下述重量比例的原料配制而成，可溶性淀粉2～3％、酵母粉0.16～0.24％、蛋白胨0.08～0.12％、氯化钠0.2～0.3％、硫酸镁0.2～0.3％、磷酸二氢钾0.04～0.06％、磷酸氢二钾0.04～0.06％、琼脂粉1.4～2.2％、余量为水。它克服了小麦品种、产地和加工等环节不稳定因素的影响，质量很难均一稳定的缺陷；它表面光滑，质量稳定均一，配方成分稳定；由此培养的菌种稳定性好，菌种菌落饱满，白色均匀，孢子同步性好，效价高，适用于菌种分离选育，提高了工作效率。
Description
技术领域
本发明属于微生物发酵工程技术领域，具体涉及一种土霉素菌种斜面培养基。
背景技术
土霉素(Oxytetracycline)是龟裂链霉菌(Streptomyces rimosusSobinetal)发酵产生的广谱抗生素。是我国生产研究早、工艺成熟、成本低、产量大的抗生素品种。在医疗、畜牧业和农业方面用途广泛。中国的土霉素发酵工艺已经达到了很高水品技术提升空间有限。而今国内企业土霉素工业生产菌种选育、制备工艺仍然使用传统的麸皮工艺，生产菌种的质量成为提高稳定发酵水平的关键。
土霉素生产菌制备工艺关键是麸皮的质量。由于麸皮的制备过程复杂，质量很难均一稳定，影响因素包括小麦品种、小麦产地、小麦加工等各环节都会严重影响麸皮的质量。
麸皮的制作经过一粒一粒的人工挑选小麦，合格的小麦在经过水洗---浸泡---控水-冷藏---研磨---过筛---洗涤---晾晒等等复杂过程，其制备过程完全凭经验控制，人为因素多、标准粗放、质量很难稳定。所以往往一个公司几个车间的麸皮差异都很明显。同时麸皮制作大大增加种子工的工作量、一般历时几个月才能彻底完成一年的麸皮储备。
发明内容
本发明的目的在于克服现有技术的缺点，提供一种质量稳定、菌种生长快、效价高的土霉素菌种斜面培养基。
本发明的目的是这样实现的：一种土霉素菌种斜面培养基，其特征在于：它由下述重量比例的原料配制而成，可溶性淀粉2～3％、酵母粉0.16～0.24％、蛋白胨0.08～0.12％、氯化钠0.2～0.3％、硫酸镁0.2～0.3％、磷酸二氢钾0.04～0.06％、磷酸氢二钾0.04～0.06％、琼脂粉1.4～2.2％、余量为水。
作为本发明的进一步改进，所述的土霉素菌种斜面培养基由下述重量比例的原料配制而成，可溶性淀粉2.25～2.75％、酵母粉0.18～0.22％、蛋白胨0.09～0.11％、氯化钠0.225～0.275％、硫酸镁0.225～0.275％、磷酸二氢钾0.045～0.055％、磷酸氢二钾0.045～0.055％、琼脂粉1.6～2.0％、余量为水。
作为本发明的更进一步改进，所述的土霉素菌种斜面培养基由下述重量比例的原料配制而成，可溶性淀粉2.5％、酵母粉0.2％、蛋白胨0.1％、氯化钠0.25％、硫酸镁0.25％、磷酸二氢钾0.05％、磷酸氢二钾0.05％、琼脂粉1.8％、余量为水。
所述的土霉素菌种斜面培养基的制备方法：
按上述比例称取可溶性淀粉、酵母粉、蛋白胨、氯化钠、硫酸镁、磷酸二氢钾和磷酸氢二钾，按比例用无离子水溶解，定容分装到预先分装好琼脂的茄瓶中；茄瓶在消毒釜经过121℃25分钟消毒，摇匀倾斜竖立放置，待培养基温度40～50℃摆放成斜面。
使用本发明土霉素菌种斜面培养基培养土霉素菌种的方法：
将土霉素菌种接种到本发明的土霉素菌种斜面培养基上，36.5±0.5℃高温培养60～70小时，然后至30±0.5℃温度继续培养20～30小时。菌种培养结束至4～6℃的冰箱保藏。
本发明的优点在于：本发明不使用小麦麸皮为培养基原料，克服了小麦品种、产地和加工等环节不稳定因素的影响，质量很难均一稳定的缺陷；本发明土霉素菌种斜面培养基表面光滑质量稳定均一，配方成分稳定、由此培养的菌种稳定性好，菌种菌落饱满，白色均匀，菌种特性没有改变，孢子同步性高，由此斜面培养基培养的菌种效价显著高于麸皮斜面培养基菌种的效价。由于菌种的稳定，为批量稳定培养生产菌种提供了良好的条件。此土霉素菌种斜面培养基表配制工艺方便易学，适用于菌种分离选育，大大地提高了工作效率。
具体实施方式
下面结合实施例进一步阐述本发明内容。
实施例1：一种土霉素菌种斜面培养基，它由下述重量比例的原料配制而成，可溶性淀粉2.5％、酵母粉0.2％、蛋白胨0.1％、氯化钠0.25％、硫酸镁0.25％、磷酸二氢钾0.05％、磷酸氢二钾0.05％、琼脂粉1.8％、余量为水。
实施例2：一种土霉素菌种斜面培养基，它由下述重量比例的原料配制而成，可溶性淀粉2.2％、酵母粉0.23％、蛋白胨0.08％、氯化钠0.3％、硫酸镁0.2％、磷酸二氢钾0.04％、磷酸氢二钾0.06％、琼脂粉1.6％、余量为水。
实施例3：一种土霉素菌种斜面培养基，它由下述重量比例的原料配制而成，可溶性淀粉2.7％、酵母粉0.18％、蛋白胨0.12％、氯化钠0.2％、硫酸镁0.3％、磷酸二氢钾0.06％、磷酸氢二钾0.04％、琼脂粉2.0％、余量为水。
实施例4：
(1)按上述实施例1的比例称取可溶性淀粉、酵母粉、蛋白胨、氯化钠、硫酸镁、磷酸二氢钾和磷酸氢二钾，按比例用无离子水溶解，定容分装到预先分装好琼脂的茄瓶中；茄瓶在消毒釜经过121℃25分钟消毒，摇匀倾斜竖立放置，待培养基温度40～50℃摆放成斜面。
(2)将土霉素菌种接种到本发明的土霉素菌种斜面培养基上，36.5±0.5℃高温培养65小时，然后至30±0.5℃温度继续培养25小时。菌种培养结束至4～6℃的冰箱保藏。
(3)将斜面培养的菌种接种于摇瓶25/250ml培养7天，比麸皮斜面培养基培养的菌种效价提高1100u/ml。
(4)将摇瓶培养的菌种接种在8吨种子罐内培养，比麸皮斜面培养基培养的菌种的种子生长快2～3小时达到进罐指标。
(5)将种子罐培养的菌种。用于27.5吨发酵罐培养160小时，比麸皮斜面培养基培养的菌种效价提高1300u/ml。
Claims (3)
1.一种土霉素菌种斜面培养基，其特征在于：它由下述重量比例的原料配制而成，可溶性淀粉2～3％、酵母粉0.16～0.24％、蛋白胨0.08～0.12％、氯化钠0.2～0.3％、硫酸镁0.2～0.3％、磷酸二氢钾0.04～0.06％、磷酸氢二钾0.04～0.06％、琼脂粉1.4～2.2％、余量为水。
2.根据权利要求1所述的土霉素菌种斜面培养基，其特征在于：它由下述重量比例的原料配制而成，可溶性淀粉2.25～2.75％、酵母粉0.18～0.22％、蛋白胨0.09～0.11％、氯化钠0.225～0.275％、硫酸镁0.225～0.275％、磷酸二氢钾0.045～0.055％、磷酸氢二钾0.045～0.055％、琼脂粉1.6～2.0％、余量为水。
3.根据权利要求2所述的土霉素菌种斜面培养基，其特征在于：它由下述重量比例的原料配制而成，可溶性淀粉2.5％、酵母粉0.2％、蛋白胨0.1％、氯化钠0.25％、硫酸镁0.25％、磷酸二氢钾0.05％、磷酸氢二钾0.05％、琼脂粉1.8％、余量为水。
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