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枯草杆菌JSIM-1019突变株肌苷发酵研究 总被引:4,自引:0,他引:4
以肌苷产生菌枯草杆菌7171-9-1为出发菌株,经物理、化学诱变剂连续处理,获得一株腺嘌呤、组氨酸、硫胺素三重营养缺陷型并对8-氮杂鸟嘌呤、6-巯基嘌呤有双重抗性的突变株JSIM-1019。在摇瓶和发酵罐试验中,该变异株的肌苷产量显著高于亲株。摇瓶试验产肌苷达20g/L,最高可达24.83g/L。工业生产试验最高达14.5g/L,稳定在12g/L。发酵周期平均为43.8小时。菌株遗传性状稳定。 相似文献
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肌苷菌核酸代谢关键酶缺失和形成选育腺苷菌的研究 总被引:3,自引:0,他引:3
以肌苷产生菌Bacillus Subtilis JSIM-1019为出发菌株,根据B.subtilis核酸代谢理论设计不同筛选模型,用物理、化学诱变剂对亲株进行诱变处理,先后有序地获得不同关键酶的缺失或回复即特殊的营养缺陷型以及抗某些代谢类似物的突变株,解除终产物对代谢物的抑制和阻遏,获得了几株黄嘌呤营养缺陷型并对8氮鸟嘌呤具有抗性的突变株X-13等,获得的突变株经单菌分离后得到X-13-4,36℃培养72h在培养基中最高积累12.43g/L腺苷。 相似文献
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该鉴定会由浙江大学陈学旺副教授主持。来自各地代表39人。枯草杆菌JSIM-1019是以7171-9-1为出发菌株,先经亚硝基胍和8-氮杂呤与处理,分离出突变株后又经紫外线和6-巯 相似文献
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已肌苷产生菌枯草杆菌GMI- 741(Ade - )为出发菌株 ,通过多因子诱变选育出具有非精确嘌呤缺陷型、丧失腺嘌呤脱氨酶的AICAR(5 -氨基 - 4-氨甲酰咪唑核苷 )产生菌AIC - 90 ,该菌株摇瓶发酵 72h产AICAR可达 2 0 .3g/L以上。 相似文献
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十六烷基三甲基溴化铵对枯草杆菌( B. subtilis )黄嘌呤缺陷型菌株产生腺苷的影响 总被引:3,自引:0,他引:3
在枯草杆菌中,嘌呤核柑酸合成途径中丰在AMP和GMP转变成IMP的环行途径,而在该菌的黄嘌呤缺陷型菌株中IMP至XMP的分枝途径被阻塞,同时在该菌株内又有强的5’一核苷酸酶活性,这使其可以生成腺苷(AR),但反馈抑制是腺苷产率的一个重要影响因素,使用季铵盐阳离子表面活性剂十六烷基三甲基溴化铵(CTAB)可改善细胞的渗透性,减少细胞内产物的反馈抑制,促进产物的分泌,结果表明,在发酵60h添加4%的CTAB能使产率从3.0g/l提高到3.6g/l,提高幅度达20%。 相似文献
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肌苷和鸟苷生产菌中嘌呤核苷合成途径三段基因序列的分析 总被引:3,自引:0,他引:3
为了研究肌苷和鸟苷生产菌中与产苷有关的嘌呤核苷合成途径的遗传背景,选择了pur操纵子的启动子序列、编码SAMP合成酶的purA基因和编码GMP合成酶的guaA基因,设计合适的引物,分别从野生菌、一株肌苷低产菌和肌苷鸟苷高产菌中扩增出相应片段,经克隆和测序后,对它们进行比较和分析。分析结果表明两株生产菌的purA基因发生了1个碱基缺失,导致阅读框发生移码突变;而鸟苷高产菌在pur操纵子的启动子部分和操纵子抑制蛋白结合区域发生了近10%的突变,可能影响整个操纵子的表达调控。 相似文献
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Yuichiro Midorikawa Takashi Akiya Akira Kuninaka Hiroshi Yoshiho 《Bioscience, biotechnology, and biochemistry》2013,77(7):1595-1598
Among various nutritional mutants with weak 5′-nucleotidase derived from Bacillus subtilis IAM 1145, the adenine-requiring mutants could convert exogenously added hypo- xanthine, guanine, xanthine and their ribosides to 5′-inosinic acid (IMP) and accumulate it in the medium. Synthesis of IMP from purine derivatives was observed predominantly in an early stage of the cultivation. The conversion was stimulated by Fe2+ or Mn2+, and markedly depressed by an excess amount of adenine in the production-medium. 相似文献
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Phosphohydrolases of a Bacillus subtilis Mutant Accumulating Inosine and Hypoxanthine 总被引:4,自引:2,他引:2 下载免费PDF全文
Although adenine-requiring auxotrophs of Bacillus subtilis accumulate large quantities of inosine or hypoxanthine, or of both, they do not accumulate inosine-5'-monophosphate (IMP). Experiments directed at understanding this phenomenon were conducted with an adenineless auxotroph and with a mutant derived from it which lacked alkaline phosphohydrolase. It was found that B. subtilis contains four different phosphohydrolases. Only one is an extracellular enzyme; it is a 5'-nucleotide phosphohydrolase which can be inhibited by addition of CuSO(4) to the medium. Of the three cellular enzymes, only one, an acid phosphohydrolase, cannot attack 5'-nucleotides; this enzyme is not repressed by inorganic phosphate. One of the two remaining surface-bound enzymes is a nonspecific alkaline phosphohydrolase which attacks both 5'-nucleotides and p-nitrophenyl phosphate; this is the only phosphohydrolase that is markedly repressed by inorganic phosphate. The other surface-bound enzyme is a nonrepressible 5'-nucleotide phosphohydrolase with double pH optima: one at neutrality and the other near pH 9.0. The experiments indicate that the absence of IMP in the extracellular broth is due to degradation of internally accumulated IMP to inosine by the cellular 5'-nucleotide phosphohydrolase. 相似文献
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An ad-9 strain of Neurospora crassa was mutagenized with ethylmethane sulfonate (5%) and selected for guanine auxotrophy. The resultant double adenine plus guanine mutant was backcrossed with wild type and a single guanine auxotroph was isolated from the progeny. In vitro assays indicated that the mutant had GMP synthetase activity comparable with wild type, but was completely lacking of IMP dehydrogenase activity. The guanine requirement can therefore be explained by the mutant's inability to convert IMP to XMP. Another guanosine auxotroph was able to adapt and grow on minimal medium after 3 days. This mutant had GMP synthetase activity comparable with wild type but had only 10% of the IMP dehydrogenase activity of wild type, which may possibly explain its ability to grow on minimal medium after 3 days. It was confirmed that the two isolates are not allelic by crossing the two and recovering 25% wild-type progeny. Out isolate must therefore be designated gua-2. 相似文献
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A biotin-requiring coryneform bacterium which produces glutamic acid was mutated to adenine dependency. The adenine-requiring strain, which excreted insoine-5′-monophosphate (IMP), was further mutated to xanthine dependency. As expected, IMP was also excreted by this mutant. The mutant strain was reverted to xanthine independence in an attempt to obtain a culture with an altered IMP dehydrogenase which would be less sensitive to feedback inhibition by guanosine-5′-monophosphate (GMP). A revertant was obtained which produced GMP and IMP, each at 0.5 g per liter. The reversion to xanthine independence had resulted in a concomitant requirement for isoleucine, leucine, and valine. Further mutation to increased nutritional requirements led to culture MB-1802, which accumulated 1 g per liter each of GMP and IMP. Both nucleotides were isolated in pure form. The concentrations of GMP and IMP produced by MB-1802 were four times that of cytidylate, uridylate, or adenylate, indicating that the mechanism of GMP and IMP production was direct and not via ribonucleic acid breakdown. 相似文献
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A. L. Demain M. Jackson R. A. Vitali D. Hendlin T. A. Jacob 《Applied and environmental microbiology》1966,14(5):821-825
A biotin-requiring coryneform bacterium which produces glutamic acid was mutated to adenine dependency. The adenine-requiring strain, which excreted insoine-5′-monophosphate (IMP), was further mutated to xanthine dependency. As expected, IMP was also excreted by this mutant. The mutant strain was reverted to xanthine independence in an attempt to obtain a culture with an altered IMP dehydrogenase which would be less sensitive to feedback inhibition by guanosine-5′-monophosphate (GMP). A revertant was obtained which produced GMP and IMP, each at 0.5 g per liter. The reversion to xanthine independence had resulted in a concomitant requirement for isoleucine, leucine, and valine. Further mutation to increased nutritional requirements led to culture MB-1802, which accumulated 1 g per liter each of GMP and IMP. Both nucleotides were isolated in pure form. The concentrations of GMP and IMP produced by MB-1802 were four times that of cytidylate, uridylate, or adenylate, indicating that the mechanism of GMP and IMP production was direct and not via ribonucleic acid breakdown. 相似文献
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Hiroshi Matsui Katsuaki Sato Hitoshi Enei Koichi Takinami 《Bioscience, biotechnology, and biochemistry》2013,77(9):2347-2352
An inosine- and guanosine-producing strain, AJ11100, of Bacillus subtilis could not grow in the minimum medium supplemented with 50 µg of sulfaguanidine per ml. When sulfaguanidine resistant mutants were derived from AJ11100, the sulfaguanidine resistance was frequently accompanied by xanthine requirement. All the xanthine auxotrophic mutants required a large amount of xanthine for cell growth and inosine accumulation. Revertants were then derived from one of the xanthine auxotrophic mutants, AJ11101, and improved inosine producers were obtained. The best mutant, AJ11102, accumulated 20.6 g of inosine per liter.Furthermore, enzyme activities of inosine 5′-monophosphate (IMP) dehydrogenase, 5′-nucleotidase and phosphoribosyl pyrophosphate (PRPP) amidotransferase were assayed to investigate why AJ11102 accumulated an increased amount of inosine. The results showed that the increase of specific activity of 5′-nucleotidase contributed much to the increased accumulation of inosine. 相似文献
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Subcellular localisation of purine-metabolising enzymes in Leishmania mexicana mexicana 总被引:1,自引:0,他引:1
H F Hassan J C Mottram G H Coombs 《Comparative biochemistry and physiology. B, Comparative biochemistry》1985,81(4):1037-1040
Leishmania mexicana mexicana cultured promastigotes were fractionated by isopycnic centrifugation on linear sucrose gradients. Guanine, hypoxanthine and xanthine phosphoribosyltransferase activities were found to be associated with glycosomes, whereas adenine phosphoribosyltransferase was cytosolic. 3'- and 5'-nucleotidases and IMP dehydrogenase were shown to be particulate, the former two possibly being associated with the plasma membrane, IMP dehydrogenase with the endoplasmic reticulum. Nucleosidases and deaminases were found to be cytosolic. The results demonstrate that intracellular separation of enzymes could play a part in the regulation of the parasite's purine metabolism. 相似文献
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Takashi Nara Masanaru Misawa Toshio Komuro Shukuo Kinoshita Kiyoshi Udagawa Shigeo Abe 《Bioscience, biotechnology, and biochemistry》2013,77(3):358-376
Enzymatic studies with Brevibacterium ammoniagenes ATCC 6872 demonstrated that 5-phosphoribose pyrophosphokinase and purinenucleotide pyrophosphorylase were involved in the nucleotide synthesis from purine base by ATCC 6872 and that its actual accumulation from base seemed to take place extracellularly through the action of the salvage enzymes leaked out of cells. Mn2+ deficiency and the simultaneous presence of pantothenate and thiamine, essential for efficient nucleotide accumulation, caused the extracellular leakage of the two enzymes with the simultaneous excretion of R5P. In the direct IMP fermentation with the adenine auxotroph, it was verified that hypoxanthine first produced de novo was reconverted into IMP extracellularly by the salvage enzymes as speculated previously.A guanine-requiring mutant of Brevibacterium ammoniagenes ATCC 6872 accumulated a large amonnt of 5′-xanthosine-monophosphate (abbreviated as XMP).The quantity of XMP accumulated by the strain was affected significantly by guanine levels in the medium. The suppression of XMP accumulation by an excessive addition of guanine compounds was recovered by the supply of casamino acids in the medium.An enzyme in the pathway of de novo XMP synthesis, IMP dehydrogenase (IMP: NAD oxidoreductase, EC 1.2.1.14), was repressed and inhibited by guanine compounds.The facts that an exogenous xanthine was not converted to XMP by the growing cells and that the activity of XMP-pyrophosphorylase was very low or deficient suggest that XMP accumulation by the strain would be probably due to the direct excretion of the nucleotide from the cells. 相似文献
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Yeast cells inhibited by benzimidazole accumulate hypoxanthine with associated efflux of xanthine. Unlike control cells, inhibited cells contain no detectable free UMP and CMP. Benzimidazole decreases uptake of [8-14C]hypoxanthine into the intracellular pool of hypoxanthine and xanthine but causes radioactive xanthine to accumulate in the medium. In inhibited cultures there is a threefold increase in incorporation of [8-14C]hypoxanthine into the total (intracellular plus extracellular) xanthine. Uptake of [8-14C]hypoxanthine into free nucleotides and into bound adenine and guanine was inhibited by 70%. Uptake of [U-14C]glycine into IMP, AMP, GMP, DNA and RNA was also substantially decreased. Incorporation of [2-14C]uracil into the intracellular uracil pool was inhibited by 30% and into free uridine and cytidine by over 90%. Benzimidazole inhibited incorporation of [8-3H]IMP into AMP and GMP, and decreased substantially the activity of glutamine-amidophosphoribosyltransferase (EC 2.4.2.14). Yeast cultures were shown to N-ribotylate benzimidazole. Results are consistent with benzimidazole inhibiting yeast growth by competing for P-rib-PP and so depriving other ribotylation processes such as the 'salvage' pathways and de novo synthesis of purines and pyrimidines. 相似文献