CN1203184C - 经微生物学方法制备假单胞菌酸a抗生素的方法 - Google Patents
经微生物学方法制备假单胞菌酸a抗生素的方法 Download PDFInfo
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- CN1203184C CN1203184C CNB008058784A CN00805878A CN1203184C CN 1203184 C CN1203184 C CN 1203184C CN B008058784 A CNB008058784 A CN B008058784A CN 00805878 A CN00805878 A CN 00805878A CN 1203184 C CN1203184 C CN 1203184C
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Abstract
公开用于制备假单胞菌酸A的方法,方法包括在通气的条件下深层培养能够生物合成假单胞菌酸A的假单胞菌菌株;且分离要求的化合物。本发明方法特别包括在大约20℃至大约30℃之间的温度下,于含有有机氮和碳源和任选含有矿物盐的培养基上,培养保藏于布达佩斯匈牙利国立农业和工业微生物收藏中心的假单胞菌菌株19/26号,保藏号为NCAIM(P)B 001235,或它的产生假单胞菌酸A突变型或变种。
Description
发明领域
本发明涉及用于制备抗生素假单胞菌酸A(Pseudomonic acid A)(莫匹罗星)的微生物学方法。
发明背景
假单胞菌酸A,也称作莫匹罗星,为一种主要对革兰氏阳性菌(如金黄色葡萄球菌(Staphylococcus aureus)、酿脓链球菌(Streptococcuspyogenes)、肺炎链球菌(Streptococcus pneumoniae)、肺炎克氏杆菌(Klebsiella pneumoniae))和一些革兰氏阴性菌(如流感嗜血杆菌(Haemophilus influenzae)、淋病奈瑟氏球菌(Neisseria gonorrhoeae))具有生长抑制作用的抗生素[A.Ward,D.M.Campoli-Richards,Drugs 32,425-444(1986)]且它的最小抑制浓度处于0.02-0.5mg/dm3范围内。通过抑制异亮氨酸-tRNA合酶,假单胞菌酸A影响病原菌的肽合成[J.Hughes和G.Mellows,Biochem.J.191,209-219(1980)]。该抗生素的一个有利特征是它对人和动物具有非常低的毒性且其在埃姆斯(Ames)试验中为阴性。目前假单胞菌酸A以多种制剂用于人的治疗,以治疗皮肤感染(例如脓疱病、脓皮病)、鼻和外耳感染、痤疮、烧伤、湿疹、牛皮癣、在溃疡情况中用于治疗二次感染,和用于预防医院感染。
假单胞菌酸A的化学结构测定如由式(I)描述的那样,为9-{4[5S(2S,3S-环氧-5S-羟基-4S-甲基己基)-3R,4R-二羟基-四氢吡喃-2S-基]-3-甲基丁-2(E)-烯酰氧基}壬酸[E.B.Chain和G.Mellows,J.C.S.Chem.Comm.847-848(1974);R.G.Alexander,J.P.Clayton,K.Luk,N.H.Rogers,T.J.King,J.C.S.Perkin I.561-565(1978)]:
已知荧光假单胞菌(Pseudomonas fluorescens)能够产生假单胞菌酸A。按英国专利1,395,907号,荧光假单胞菌NCIB 10586菌株能够生物合成由假单胞菌酸A和它的在C2和C3碳原子之间的双键处于顺式位置的异构体和假单胞菌酸B组成的假单胞菌酸复合体。成分比例为4.5∶4.5∶1。然而,按照日本专利申请52-70083号,荧光假单胞菌Y-11633菌株能够生物合成由以9∶0.5∶0.5的比例存在的假单胞菌酸A、假单胞菌酸B和另外两种具有未知结构的成分组成的假单胞菌酸复合体。
发明概述
本发明涉及制备假单胞菌酸A的方法,方法包括:在深层通气的条件下,于包含至少一种有机氮或碳源的培养基上培养能够生物合成假单胞菌酸A的假单胞菌属菌株,且发酵假单胞菌属菌株培养物以便形成假单胞菌酸A。假单胞菌菌株优选为匈牙利布达佩斯国立农业和工业微生物保藏中心(National Collection of Agricultural andIndustrial Microorganisms)保藏的假单胞菌菌株(Pseudomonas sp.Bacterium strain)19/26号,保藏号NCAIM(P)B 001235,或它的产生假单胞菌酸A的突变型或变种。
本发明也涉及在深层通气的条件下能够生物合成假单胞菌酸A的假单胞菌培养基,其基本上由新的假单胞菌菌株19/26号组成。
本发明另外涉及新的假单胞菌菌株19/26号的生物纯的培养基。
图的简述
图1为由假单胞菌菌株19/26号(NCAIM(P)B 001235)产生的假单胞菌酸复合体的HPLC色谱图。
发明详细描述
在寻找细菌产生的抗微生物抗生素过程中,在含有为预防真菌生长的10μg/ml对羟苯乙基三甲基铵和5μg/ml环己酰亚胺的营养琼脂培养基上,分离到20,000种微生物。
在不同的革兰氏阳性和革兰氏阴性试验微生物上,检查所分离的菌株的摇瓶培养基的抗菌有效性和它们对治疗实践中出现的对抗生素抵抗的变种。通过应用抗生素抵抗的试验微生物,我们想促进识别具有新的作用模式的抗菌抗生素。
在上述的检查过程中,选择了称作“菌株19/26号”的细菌分离物。最初,已从阿根廷收集的土壤样品得到该微生物。其培养液分别对枯草酸杆菌(Bacillus subtilis)ATCC 6633、金黄色葡萄球菌SMITH、耐青霉素金黄色葡萄球菌1110、未编号的肺炎链球菌菌株、化脓链球菌A 115 ROBB、粪产碱杆菌(Alcaligenes faecalis)140001、支气管败血性博代氏杆菌(Bordetella bronchiseptica)ATCC 4617、克雷伯氏肺炎杆菌ATCC10031、耐美他西林和耐氨基糖苷类的金黄色葡萄球菌MRSA 1190R和两种未编号的肺炎支原体(Mycoplasmapneumoniae)和流感嗜血杆菌菌株具有明显的抗菌作用。之后,从菌株19/26号的培养液分离到其抗菌代谢产物,且化学结构分析结果发现与假单胞菌酸A一致。
同时,对选择的产生假单胞菌酸A的菌株进行分类研究。首先,使用装配ID 32 GN条的Bio-Merieux ATB表达,用选择的密切相关的菌株进行初步比较研究。后者适宜于相对快(如属的水平)的所给出的未知菌株的分类鉴定方法,如果它的诊断性质能够显示出所需的与至少一种这些被确认的菌株的高度表型相似性,这些被确认的菌株加入在设备的数据库中。用这个设备分别对14种不同的糖和14种有机酸的32次试验,能够测定所研究的未知菌株碳源利用图谱,加入到一种用生长因子复合体完全的基本培养基中。在接种和经过48小时孵育时间后,借助自动浊度测量法,在标明生长强度(反应)分别为+或-或?的发育微量培养上,进行利用试验结果评价。通过设备软件,评价同步研究的生物化学试验结果。所得到的菌株19/26号的生理学-生物化学数据与数据库中列出的菌株数据相一致(在该情况中,这样的菌株总数为114种,其中有14种属于不同的假单胞菌种类:铜绿假单胞菌(Pseudomonas aeruginosa)I和II、产碱假单胞菌(Pseudomonas alcaligenes)、荧光假单胞菌I和II、门多萨假单胞菌(Pseudomonas mendocina)、嗜温假单胞菌(Pseudomonas mesophilica)、皮氏假单胞菌(Pseudomonas rpickettii)、类鼻疽假单胞菌(Pseudomonaspseudomallei)、泡囊假单胞菌(Pseudomonas vesicularis)、洋葱假单胞菌(Pseudomonas cepacia)、缺陷假单胞菌(Pseudomonas diminuta)、恶臭假单胞菌(Pseudomonas putida)和旋氏假单胞菌(Pseudomonasstutzeri))。按照这个相关实验的结果,证实菌株19/26号为假单胞菌属的一个成员。
在菌株19/26号的属的水平上成功鉴定后,我们尝试借助经典分类研究方法在种的水平上鉴定它。
在以非孢子形成的革兰氏阴性杆菌的形式存在的适宜的固体培养基上,菌株19/26号的培养基能够增加0.6-1.1至1.3-4.0微米。这些是靠极性鞭毛主动游动的。这些专性需氧的化能有机营养杆菌仅能够氧化侵蚀(attack)葡萄糖。它们不能发酵且不能用硝酸盐作为末端电子受体被修复,但显示阳性催化酶反应。它们在简单的合成培养基上不能发展可见菌落且对它们的增殖总是需要生长因子。在最佳条件下生长,它们的菌落呈精氨酸二水解酶阳性,产生荧光色素但在细胞中不能检测多-β-羟基-丁酸盐的存在和聚集。
菌株19/26号的其它特征如下:在41℃下不能观察到它的菌落发展;它从蔗糖中不产生果聚糖,不水解明胶和淀粉,其呈氧化酶阳性,且不利用葡萄糖、2-酮基-葡糖酸盐、海藻糖和内消旋肌醇作为单一碳源。
所有这些诊断性质将假单胞菌菌株19/26号与以下菌种清楚地分开:铜绿假单胞菌、荧光假单胞菌、绿叶假单胞菌、致金色假单胞菌、丁香假单胞菌、绿黄假单胞菌、菊苣假单胞菌、旋氏假单胞菌、门多萨假单胞菌、产碱假单胞菌、类产碱假单胞菌和恶臭假单胞菌。菌株19/26号能够仅在用(0.2-1.0%)酵母提取物、玉米浸渍液或其它的含有不同种类生长因子的复合天然营养物制备的培养基上进行培养。
在这些特殊诊断性质的基础上,认为菌株19/26号能够属于所谓“非典型假单胞菌种”有机体。这样的“非典型假单胞菌种”未来可加入到新建立的细菌分类群中(N.J.Palleroni,“Psuedomonaceae”Bergeys Manual of Systematic Bacteriology,1,141-219,N.R.Krieg和J.G.Holt编辑,Baltimore:Williams and Wilkins,1984)。
菌株19/26号于1996年7月16日保藏于匈牙利布达佩斯国立农业和工业微生物保藏中心,保藏号为NCAIM(P)B 001235。
按照本发明优选方法,以保藏号NCAIM(P)B 001235保藏的假单胞菌属菌株19/26号用于制备基本纯的假单胞菌酸A。所选择的菌株能够使用蛋白胨、牛肉膏、玉米浸渍液、酵母提取物、酪蛋白和大豆粉作为氮源。除以上氮源以外,碳源例如葡萄糖、甘油和葵花油能够以不同的组合使用。
尽管天然来源的培养基成分含有无机盐,向接种培养基中加入一些无机盐(例如铵、钙、铁、锌、铜、镁、锰、钠或钾盐)仍是有利的。优选硫酸镁、二氯化锰、硫酸亚铁、氯化锌、硫酸铜II、硫酸铵、磷酸二氢钾、氯化钠和碳酸钙。
在4℃下,在含有蛋白胨-酪蛋白的斜面琼脂培养基上进行菌株的维持,每两周将它新鲜转移。为维持假单胞菌酸A的生产力,菌株能够经深度冷冻培养基或以冷冻干燥形式适宜贮存。
在发酵期间,将假单胞菌种19/26号菌株接种到适宜的培养基上并在半浸渍和通气发酵条件下培养。培养基的pH优选设置在中性值(pH=大约7.0),培养温度在大约20℃至大约30℃之间,优选在大约24℃至大约26℃之间。依发酵条件而定,50-60小时后能够达到抗生素生产力的最大值。在发酵期间形成的假单胞菌酸复合体包含基本纯的假单胞菌酸A,尽管作为生物合成的结果,也形成了少量的式(II)假单胞菌酸B和式(III)假单胞菌酸C。
经琼脂扩散微生物方法,测定假单胞菌酸抗菌复合体的抗菌活性。培养基为含有牛肉膏-蛋白胨-葡萄糖的琼脂,其pH为6.5且试验有机体为枯草杆菌ATCC 6633。经微生物学方法得到的活性值主要表示假单胞菌酸A的量,因为其它假单胞菌酸成分的量非常小,且相对于成分A,它们对试验-微生物的特异活性-尤其是在成分B的情况下-被认为更弱。
在发酵期间,通过高效液相色谱法(HPLC)测定在发酵液体培养基中准确量的假单胞菌酸A和伴随的次要成分,其中经乙醇稀释两次的液体培养基的超声处理和离心的样品的上清液被研究(装置:LKB2248泵、LKB 2141 UV检测器(在222nm下分析),柱:Nucleosil C8 10μm(BST),洗脱液:乙腈和0.1M NH4H2PO4溶液(pH:5.0)的混合物(35∶65),流速:1.2ml/min),保留时间:假单胞菌酸A(PSA)为8.5min,假单胞菌酸B(PSB)为6.0min,且假单胞菌酸C(PSC)为22.5min。
在图1中能够看到经假单胞菌属19/26号菌株生物合成的假单胞菌酸抗菌复合体的HPLC色谱图。该色谱图表明经基本纯的假单胞菌酸A的发酵酸19/26号菌株得到假单胞菌酸复合体。假单胞菌酸B和假单胞菌酸C成分的总量低于5%。以这种方式证明假单胞菌属19/26号菌株能够以比早先公开的产生假单胞菌酸复合体的荧光假单胞菌菌株以更优良的组合来生物合成抗生素。这个事实对工业生产是有利的。
通过以下实施例阐明本发明的方法。然而,本发明应不受其限制。
实施例
在PCA标记的琼脂斜面培养基上维持假单胞菌属19/26号菌株。
PCA培养基的组分如下:
牛肉膏 3g
蛋白胨 5g
琼脂 15g
用蒸馏水稀释至1000ml。培养基的pH为7.0-7.2。
在25℃下,将接种的琼脂斜面培养基孵育24小时,并把细胞悬浮于5ml生理盐水溶液(悬浮液中的细胞数:109-1010/ml)中。在500ml锥形瓶中,将得到的1ml悬浮液接种到灭菌的100ml的I-21标记的接种培养基中。I-21培养基组分如下:
葡萄糖 10g
甘油 5g
玉米浸渍液 3g
硫酸铵 2g
磷酸二氢钾 0.4g
硫酸镁-水(1∶7) 0.4g
二氯化镁-水(1∶2) 0.03g
碳酸钙 4g
葵花油 2g
用自来水稀释至1000ml。灭菌前培养基的pH设定为7.0。
在25℃下,于摇床(260RPM,振幅10cm)上将含有移种培养基的烧瓶振摇18-20小时。之后,将50ml(1%)的振摇培养基接种到E-5标记的5升培养液中,在121℃下,于10升夹套发酵罐中灭菌45分钟。E-5培养基的组分如下:
葡萄糖* 50g
甘油 50g
大豆粉 100g
玉米浸渍液 15g
氯化钠 25g
碳酸钙 25g
葵花油 10g
用自来水稀释至5升(*葡萄糖在50%溶液中灭菌30分钟,并与接种物质一起加入到培养基中)。灭菌前培养基的pH设定为7.0。
搅拌培养液并通气50-60小时。培养液的温度为25℃,搅拌速率为500RPM,空气流速为200升/小时。以最小量(每发酵罐大约20-30ml)使用作为抗泡沫剂的葵花油。
假单胞菌酸复合体的生物合成在发酵8-10小时时开始且在培养55-60小时时实验最大抗菌浓度。
如下进行从发酵液体培养基中分离假单胞菌酸A。在完成发酵后,将得到的4.5升培养发酵液离心,然后用稀(20%)硫酸将上清液(4.06升)的pH调至4.5。然后用2.03升乙酸乙酯将酸化液提取两次。分离这些相并经离心从乳液状有机相制备清晰的相。真空蒸发合并的提取液。将得到的粗产物(2.45g)溶解于25ml氯仿-甲醇-99.5%乙酸(93∶5∶2)的混合液中,且将得到的溶液填充到从245g的Kieselgel60(粒子大小:0.063-0.2mm;Reanal)和用以上溶剂混合物制备的柱(高∶直径=28∶5)上。用以上溶剂混合物洗脱。在洗脱过程中,收集50ml流分并经薄层层析法分析流分中假单胞菌酸A的含量,使用Kieselgel60(DC-Alufolien;105554,默克)吸附剂和氯仿-甲醇-99.5%乙酸(90∶8∶2)展开溶剂混合物。从Kieselgel 60柱上洗脱的流分34-50包含假单胞菌酸A。合并这些馏分(850ml)并同时将冷却的280ml水加入到得到的溶液中。在此之后,用1N氢氧化钠水溶液将溶液混合物的pH调至4.5。从水相中分离有机溶液,然后经280ml氯仿再次提取水相。真空蒸发合并的提取液,如此能够得到纯的假单胞菌酸A。
柱色谱馏分14-15含有来自次要成分的假单胞菌酸C,而馏分54含有来自次要成分的假单胞菌酸B,通过以上写出的方法能够以纯的形式回收它们。
使用以上显示的结构式(I)的位次编号系统,描述所分离的假单胞菌酸成分的光谱特征。
假单胞菌酸A的光谱特征:
紫外光谱(10μg/ml,在95%乙醇溶液中):λmax=222nm
红外光谱(KBr):νOH 3483和3306,νC=O 1728(COOCH2),1720(COOH)cm-1
1H-NMR谱(CDCl3,δTMS=0):
δ[ppm], 偶合常数(Hz) 归属
(积分),峰裂数
5.75(1H)q 4J2,15=1.1 2-H
4.08(2H)t 3J8',9'=6.4 9′-H2
3.72-3.93(4H)m 5-H;7-H;13-H;16-Ha
3.55(1H)dd 2J16a,16b=11.8;3J16b,8=2.6 16-Hb
3.48(1H)dd 3J5,6=8.4;3J6,7=3.2 6-H
2.82(1H)td 3J9,10=6.3;3J10,11=2.3 10-H
2.74(1H)dd 3J10,11=2.3;3J11,12=7.8 11-H
2.60(1H)dd 2J4a,4b=14.5;3J4a,5=2.7 4-Ha
2.28-2.36(3H)m 4-Hb;2′-H2
2.20(3H)d 4J2,15=1.1 15-H3
2.02(1H)m 8-H
1.61-1.76(6H)m 9-H2;3′-H2;8′-H2
1.33-1.43(9H)m 12-H;4′-H2;5′-H2′;6′-H2;7′-
H2
1.22(3H)d 3J13,14=6.4 14-H3
0.94(3H)d 3J12,17=7.0 17-H3
13C-NMR谱(CDCl3溶液,δTMS=0):
δ[ppm] 归属 δ[ppm] 归属
177.8s C-1′ 42.7t,d C-4,C-12
166.9s C-1 39.4d C-8
156.0s C-3 33.9t,t C-9,C-2′
117.7d C-2 31.6t C-4′*
74.9d C-5 28.9t C-5′*
71.4d C-13 28.8t C-6′*
70.4d C-7 28.5t C-8′*
69.0d C-6 25.9t C-7′
65.3t C-16 24.6t C-3′
63.9t C-9′ 20.8q C-14
61.3d C-11 19.1q C-15
55.6d C-10 12.7q C-17
*可交换归属
化学电离(CI)质谱:
特征谱数据:
m/z R.I.(%) 归属
501 100 [M+H]+
327 45 [M+H-HO/CH2/8COOH]+
309 16 [m/z 327-H2O]+
227 33 [C12H19O4]+
假单胞菌酸B的光谱特征:
紫外光谱(10μg/ml,在95%乙醇溶液中):λmax=222nm
红外光谱(薄膜):νOH3418,νC=O 1713(COOCH2,COOH)cm-1
1H-NMR谱(CDCl3,δTMS=0):
δ[ppm], 偶合常数(Hz) 归属
(积分),峰裂数
5.68(1H)s 2-H
4.02(2H)t 3J8',9'=6.6 9′-H2
3.7(2H)m 7-H,13-H
3.55(1H)td 3J4a,5=3J5,6=9.3;3J4b,5=1.8 5-H
3.41(2H)dd 2J16a,16b=11.0 16-H2
3.24(1H)dd 3J5,6=9.3;3J6,7=2.8 6-H
2.92(1H)td 3J9,10=5.6;3J10,11=2.0 10-H
2.69(1H)dd 3J10,11= 2.0;3J11,12=7.2 11-H
2.62(1H)dd 2J4a,4b=14.5;3J4a,5=1.9 4-Ha
2.20(2H)t 3J2',3'=7.4 2′-H2
2.14(3H)br.s 15-H3
2.10(1H)dd 2J4a,4b=14.5;3J4b,5=9.3 4-Hb
1.85(1H)dd 2J9a,9b=14.3;3J9a,10=5.3 9-Ha
1.20-1.68(14H)m 9-Hb;12-H;3′-H2:4′-H2;5′-
H2;6′-H2;7′-H2;8′-H2
1.14(3H)d 3J13,14=6.4 14-H3
0.88(3H)d 3J12,17=7.1 17-H3
化学电离(CI)质谱:
特征谱数据:
m/z RI(%) 归属
517 100 [M+H]+
343 70 [M+H-HO/CH2/8COOH]+
假单胞菌酸C的光谱特征:
紫外光谱(10μg/ml,在95%乙醇溶液中):λmax=222nm
红外光谱(薄膜):νOH 3435,νC=O 1713(COOCH2,COOH)cm-1
1H-NMR谱(CDCl3,δTMS=0):
δ[ppm], 偶合常数(Hz) 归属
(积分),峰裂数
5.69(1H)s 2-H
5.25-5.50(2H)m 10-H;11-H
4.00(2H)t 3J8',9′=6.5 9′-H2
3.88(1H)dd 3J6,7=3J7,8=3.0 7-H
3.78(1H)dd 2J16a,16b=11.8;3J16a,8=2.6 16-Ha
3.68(1H)dd 3J4b,5=2.5;3J5,6=9.0 5-H
3.55(1H)qd 3J12,13=3J13,14=6.3 13-H
3.49(1H)dd 2J16a,16b=11.8;3J16b,8=2.0 16-Hb
3.41(1H)dd 3J5,6=9.0;3J6,7=3.0 6-H
2.58(1H)dd 2J4a,4b=13.8;3J4a,5=2.5 4-Ha
2.20(2H)t 3J2′,3′=7.4 2′-H2
2.00-2.30(4H)m 4-Hb;9-H2;12-H
2.10(3H)m 15-H3
1.78(1H)m 8-H
1.45-1.65(4H)m 3′-H2;8′-H2
1.15-1.35(8H)m 4′-H2;5′-H2;6′-H2,7′-H2
1.08(3H)d 3J13,14=6.3 14-H3
0.92(3H)d 3J12,17=6.8 17-H3
化学电离(CI)质谱:
特征谱数据:
m/z R.I.(%) 归属
485 30 [M+H]-
287 60 [M+H-HO/CH2/8COOH]′
尽管在此已描述本发明一些目前优选的实施方案,对本发明从属的领域技术人员显而易见的是可进行所描述的实施方案的变化和改进,而不背离本发明的精神和范围。因此,本发明打算仅由所附的权利要求需要的内容和法律应用原则来限制。
Claims (9)
1.一种用于制备假单胞菌酸A的方法,包括以下步骤
在深层通气的条件下,于包含至少一种有机氮或碳源的培养基上培养能够生物合成假单胞菌酸A的假单胞菌属菌株,和
发酵假单胞菌属菌株培养物以便形成假单胞菌酸A,
其中所述假单胞菌属菌株为保藏于匈牙利布达佩斯国立农业和工业微生物保藏中心的假单胞菌属菌株(Pseudomonas sp.)19/26号,保藏号为NCAIM(P)B 001235。
2.权利要求1的方法,该方法还包括分离假单胞菌酸A的步骤。
3.权利要求1的方法,其中所述至少一种有机氮或碳源选自葡萄糖、甘油、葵花油、酪蛋白、大豆粉、蛋白胨、牛肉膏、玉米浸渍液和酵母提取物。
4.权利要求1的方法,其中所述培养基包含至少一种无机盐。
5.权利要求4的方法,其中所述至少一种无机盐选自铵、钙、铁、锌、铜、镁、锰、钠和钾盐和它们的混合物。
6.权利要求5的方法,其中所述至少一种无机盐选自硫酸铵、碳酸钙、硫酸亚铁、氯化锌、硫酸铜II、硫酸镁、二氯化锰、氯化钠和磷酸二氢钾和它们的混合物。
7.权利要求1的方法,其中所述发酵在大约20℃至大约30℃之间的温度下进行。
8.权利要求7的方法,其中所述发酵在大约24℃至大约26℃之间的温度下进行。
9.一种假单胞菌属菌株19/26号的生物纯培养物,该菌株保藏于匈牙利布达佩斯国立农业和工业微生物保藏中心,保藏号为NCAIM(P)B 001235。
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CN108277243B (zh) * | 2018-01-19 | 2021-03-12 | 浙江瑞邦药业股份有限公司 | 一种假单胞菌酸a的制备方法 |
CN108949845B (zh) * | 2018-08-08 | 2021-08-10 | 福建康鸿生物科技有限公司 | 一种发酵培养基及由发酵培养基制备莫匹罗星的方法 |
IL305014A (en) | 2021-02-07 | 2023-10-01 | Hangzhou Zhongmeihuadong Pharmaceutical Co Ltd | Fermentation method for mupirocin |
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JPS5270083A (en) * | 1975-12-04 | 1977-06-10 | Yoshitomi Pharmaceut Ind Ltd | Manufacture of an antibiotics, trans-pseudomonic acid |
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GB8428952D0 (en) | 1984-11-16 | 1984-12-27 | Beecham Group Plc | Formulations |
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KR20000048812A (ko) | 1996-10-01 | 2000-07-25 | 스튜어트 알. 수터, 스티븐 베네티아너, 피터 존 기딩스 | 병원성 유기체에 의한 비인두 전이증식 관련 세균 감염 치료용약제를 제조하기 위한 뮤피로신의 용도 |
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BR0009179A (pt) | 1999-02-03 | 2001-11-27 | Biogal Gyogyszergyar | Processo de preparação do antibiótico ácidopseudomÈnico a pelo método microbiológico |
HUP0105286A3 (en) | 1999-02-03 | 2003-03-28 | Biogal Gyogyszergyar | Process for the isolation of pseudomonic acid a from pseudomonic acid complex-containing culture broth |
US6335023B1 (en) | 1999-06-30 | 2002-01-01 | Ruey J. Yu | Oligosaccharide aldonic acids and their topical use |
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2002
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MXPA01007806A (es) | 2003-06-04 |
CZ20012724A3 (cs) | 2001-12-12 |
WO2000046389A1 (en) | 2000-08-10 |
YU55601A (sh) | 2003-02-28 |
SK10892001A3 (sk) | 2002-06-04 |
EP1151131A1 (en) | 2001-11-07 |
CN1670217A (zh) | 2005-09-21 |
HK1041903A1 (zh) | 2002-07-26 |
AU2980800A (en) | 2000-08-25 |
AU762768B2 (en) | 2003-07-03 |
US20030100083A1 (en) | 2003-05-29 |
BR0009179A (pt) | 2001-11-27 |
EP1151131A4 (en) | 2004-05-12 |
CN1354798A (zh) | 2002-06-19 |
IL144679A0 (en) | 2002-06-30 |
IS6028A (is) | 2001-07-31 |
HRP20010576A2 (en) | 2002-08-31 |
US7078223B2 (en) | 2006-07-18 |
JP2003515311A (ja) | 2003-05-07 |
BG105776A (en) | 2002-05-31 |
US20020042103A1 (en) | 2002-04-11 |
NO20013791D0 (no) | 2001-08-02 |
US6509177B1 (en) | 2003-01-21 |
KR20010113674A (ko) | 2001-12-28 |
TR200102234T2 (tr) | 2002-10-21 |
PL358615A1 (en) | 2004-08-09 |
IL144679A (en) | 2007-06-17 |
CA2360865A1 (en) | 2000-08-10 |
US6506591B2 (en) | 2003-01-14 |
NO20013791L (no) | 2001-10-02 |
WO2000046389A9 (en) | 2001-09-07 |
HUP0300363A2 (hu) | 2004-06-28 |
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