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1.
Bacillus licheniformis ATCC 9945A was grown on Medium E in batch fermentations in which the pH was maintained at 5.5., 6.5, 7.4, and 8.25. The effects of pH on cell growth, carbon source utilization, and gamma-polyglutamic acid (gamma-PGA) production, molecular weight, and polymer stereochemistry were determined. The gamma-PGA yield was highest (15 g/L, 96 h growth time) at pH 6.5. The increase in gamma-PGA formation at pH 6.5 corresponded with a relatively high specific production rate at high gamma-PGA concentration (0.09 h(-1), approximately 15 g/L gamma-PGA). In contrast, the specific gamma-PGA production rates at fermentor pH values of 5.5 and 7.4 decreased significantly for gamma-PGA fermentor yields > approximately 5 g/L. Interestingly, alteration of the medium pH had little to no significant effects on the product quality as measured by stereochemical composition and molecular weight. While glutamate and glycerol utilization were similar as a function of pH, citrate consumption increased at pH 6.5, indicating that the formation of gamma-PGA from citrate at pH 6.5 was of increased importance. The effect of aeration was evaluated by increasing the agitation speed (250 to 800 rpm) and aeration rate (0.5 to 2.0 L/min) at pH 6.5, the pH of maximal gamma-PGA production. Increased aeration resulted in doubling of the cell dry weights (2 to 4 g/L), increasing gamma-PGA yields (6.3 to 23 g/L by 48 h) and increasing in the maximum gamma-PGA-specific production rate (0.09 to 0.11 h(-1)). Other effects of increased agitation included a rapid depletion of glutamate and citrate (by 50 h) and a decrease in product molecular weight. Despite the increase in agitation and aeration, oxygen limitation of the culture was not avoided, because the partial pressure decreased to <1.0% by 29 h. (c) 1996 John Wiley & Sons, Inc.  相似文献   

2.
We investigated the enhancement of bioethanol production in Enterobacter aerogenes ATCC 29007 by co-fermentation of carbon sources such as glycerol, glucose, galactose, sucrose, fructose, xylose, starch, mannitol and citric acid. Biofuel production increases with increasing growth rate of microorganisms; that is why we investigated the optimal growth rate of E. aerogenes ATCC 29007, using mixtures of different carbon sources with glycerol. E. aerogenes ATCC 29007 was incubated in media containing each carbon source and glycerol; growth rate and bioethanol production improved in all cases compared to those in medium containing glycerol alone. The growth rate and bioethanol production were highest with mannitol. Fermentation was carried out at 37 °C for 18 h, pH 7, using 50 mL defined production medium in 100 mL serum bottles at 200 rpm. Bioethanol production under optimized conditions in medium containing 16 g/L mannitol and 20 g/L glycerol increased sixfold (32.10 g/L) than that containing glycerol alone (5.23 g/L) as the carbon source in anaerobic conditions. Similarly, bioethanol production using free cells in continuous co-fermentation also improved (27.28 g/L) when 90.37 % of 16 g/L mannitol and 67.15 % of 20 g/L glycerol were used. Although naturally existing or engineered microorganisms can ferment mixed sugars sequentially, the preferential utilization of glucose to non-glucose sugars often results in lower overall yield and productivity of ethanol. Here, we present new findings in E. aerogenes ATCC 29007 that can be used to improve bioethanol production by simultaneous co-fermentation of glycerol and mannitol.  相似文献   

3.
Tween-80, dimethyl sulfoxide (DMSO), and glycerol could be used as novel materials to regulate the central carbon metabolic pathway and improve gamma-PGA biosynthesis by Bacillus subtilis CGMCC 0833. With glycerol in the medium, the activity of 2-oxoglutarate dehydrogenase complex at the key node of 2-oxoglutarate was depressed, more carbon flux distribution was directed to synthesize glutamate, the substrate of gamma-PGA, which led to overproducing of gamma-PGA, reached 31.7 g/l, compared to the original value of 26.7 g/l. When Tween-80 or DMSO was in the medium, the activity of isocitrate dehydrogenase was stimulated, the branch flux from 2-oxoglutarate to glutamate was also enhanced due to the increasing of total flux from iso-citrate to 2-oxoglutarate, then a large amount of glutamate was produced, and formation of gamma-PGA was also improved, which was a different process compared with that of glycerol. Moreover, with the addition of Tween-80 or DMSO, cell membrane permeability was increased, which facilitated the uptake of extracellular substrates and the secretion of gamma-PGA by this strain; therefore, gamma-PGA production was further stimulated, and 34.4 and 32.7 g/l gamma-PGA were obtained, respectively. This work firstly employed additives to improve the biosynthesis of gamma-PGA and would be helpful in understanding the biosynthesis mechanism of gamma-PGA by Bacillus species deeply.  相似文献   

4.
The relationship between tricarboxylic acid (TCA) and glyoxalate cycle and the effect of their metabolites levels on the vancomycin production of Amycolatopsis orientalis were investigated in different concentration of glycerol (2.5-20 g/l). Intracellular glycerol levels increased with respect to increases in glycerol concentrations of the growth medium. Extracellular glycerol levels decreased slowly up to 24 h while uptake rates were increased during 36-48 h for 10 and 15 g/l and during 36-60 h at 20 g/l of glycerol. Intracellular citrate, alpha-ketoglutarate, fumarate levels increased up to 10 g/l glycerol concentration. However, intracellular succinate and malate levels were increased up to 15 g/l glycerol. Extracellular citrate, alpha-ketoglutarate, succinate and malate levels increased with respect to increases in glycerol concentration. The highest alpha-ketoglutarate dehydrogenase activity was determined at 15 g/l glycerol. Isocitrate lyase activity showed a positive correlation with the increases in glycerol concentration of the growth medium. Vancomycin production increased with the increases in glycerol concentration from 5 to 10 g/l. These results showed that A. orientalis grown in glycerol containing medium used glyoxalate shunt actively instead of TCA cycle which supports precursors of many amino acid which are effective on the antibiotic production.  相似文献   

5.
During the production of biodiesel, a significant amount of glycerol is generated which currently has little commercial value. A study on the growth and lipid production of Chlorella protothecoides using glycerol as the carbon source was performed to demonstrate the utility of recycling crude glycerol created during biodiesel production. Glycerol was examined as both the sole carbon source and in combination with glucose. Algae cultures grown on only glycerol in shake flasks showed a specific growth rate and final lipid yield of 0.1/h and 0.31 g lipid/g substrate, respectively. The values were similar to those observed on pure glucose, 0.096/h and 0.24 g lipid/g substrate. When the media contained a mixture of glycerol and glucose, simultaneous uptake of the two substrates was observed. Due to the difference in rates of lipid storage, lipid production was 0.077 g lipid/(l h) during growth on glycerol, while growth on glucose had a productivity of 0.096 g lipid/(l h). During growth on the 9:1 mixture of both glucose and glycerol, lipid productivity was 0.098 g lipid/(l h). In order to simulate the use of waste glycerol from biodiesel production the experiments were repeated and similar growth rates, yields, and lipid productivities were achieved. Therefore, we have demonstrated the promise for simultaneous high growth rates and lipid yields of C. protothecoides heterotrophically grown on mixtures of glycerol.  相似文献   

6.
Corynebacterium glutamicum ATCC 17965 was cultivated in a 4-L batch aerated fermentor with glucose, fructose and mixtures of these two sugars in various proportions as carbon sources and with different concentrations of minerals and vitamins. A multilayer centrifugation technique was devised to obtain cell extracts in order to assess intracellular production of glutamate and partitioning between intracellular and extracellular spaces for lactate and acetate, the main by-products produced during the growth phase. Glutamate production increased with the proportion of glucose in the carbon source. The average value for the intracellular concentration of glutamate obtained with basic glucose medium was increased three-fold when initial concentrations of vitamins and minerals were increased four-fold. In this case, overall production of glutamate (16.3 mM) reached the highest value obtained. Production of acetate was weak on all media types (< 1.6 mm). it was the same for lactate synthesis in media where glucose remained the major carbon source (< 2.3 mm). production of lactate was significantly higher on media where fructose was the main carbon source (> 10 mM to 60 mM). The increase in lactate production and the decrease in glutamate production were correlated to a modification of carbon flux distribution between the metabolic pathways as the fructose proportion was increased. An increase in the concentration of minerals favoured production of glutamate during growth. This was correlated with an increase in the NADPH,H+ production rate. Received 16 January 1996/ Accepted in revised form 14 January 1997  相似文献   

7.
In this study, crude glycerol with high potassium concentration was purified using acid treatment and used as carbon source for lipid production using Yarrowia lipolytica SKY7. The crude glycerol was purified using phosphoric acid (pH 2) followed by centrifugation. When purified glycerol was used as carbon source for fermentation, higher biomass productivity (0.54 g/L/h) and lipid productivity (0.2 g/L/h) was observed at 96 h compared to crude glycerol. Results indicated that 6.32 g/L potassium in crude glycerol medium was inhibitory for cell growth and lipid production by Y. lipolytica. Yield coefficients, productivities and specific growth rates were calculated for each glycerol medium. The process performance with purified glycerol medium was comparable to that of pure glycerol medium. A higher lipid yield was obtained in purified glycerol medium (0.21 g/g glycerol) than crude glycerol medium (0.124 g/g glycerol). During purification of crude glycerol, KH2PO4 was also produced as by-product. This study provides a way for valorization of crude glycerol with high potassium concentration for microbial lipid production.  相似文献   

8.
The relationship between tricarboxylic acid (TCA) and glyoxalate cycle and the effect of their metabolites levels on the vancomycin production of Amycolatopsis orientalis were investigated in different concentration of glycerol (2.5–20 g/l). Intracellular glycerol levels increased with respect to increases in glycerol concentrations of the growth medium. Extracellular glycerol levels decreased slowly up to 24 h while uptake rates were increased during 36–48th h for 10 and 15 g/l and during 36–60th h at 20 g/l of glycerol. Intracellular citrate, α-ketoglutarate, fumarate levels increased up to 10 g/l glycerol concentration. However, intracellular succinate and malate levels were increased up to 15 g/l glycerol. Extracellular citrate, α-ketoglutarate, succinate and malate levels increased with respect to increases in glycerol concentration. The highest α-ketoglutarate dehydrogenase activity was determined at 15 g/l glycerol. Isocitrate lyase activity showed a positive correlation with the increases in glycerol concentration of the growth medium. Vancomycin production increased with the increases in glycerol concentration from 5 to 10 g/l. These results showed that A. orientalis grown in glycerol containing medium used glyoxalate shunt actively instead of TCA cycle which supports precursors of many amino acid which are effective on the antibiotic production.  相似文献   

9.
以一株表达人胰高血糖素样肽-1融合蛋白的重组大肠杆菌为研究对象,首先通过摇瓶实验对碳源种类进行了初步选择,发现葡萄糖和甘油对菌体生长以及GLP-1融合蛋白表达较为适宜。进一步在5 L反应器上对初始葡萄糖及甘油浓度进行了考察,发现高浓度碳源有利于菌体生长却抑制GLP-1融合蛋白表达,但能提高GLP-1融合蛋白的体积得率。在0.25%初始葡萄糖或甘油存在的条件下,在培养过程中流加葡萄糖或甘油维持其在发酵液中的浓度,比较了两者对菌体生长以及GLP-1融合蛋白表达的影响,结果发现,以甘油为碳源时,菌体生长以及GLP-1融合蛋白的表达量均高于以葡萄糖为碳源的结果,最终发酵液的菌浓(OD_(600))可达到25.4,较葡萄糖为碳源时19.1提高了33.0%,GLP-1融合蛋白表达水平和体积得率分别可达到22.4%和1.051 g/L,较葡萄糖为碳源的15.8%和0.504 g/L分别提高41.8%和108.5%。该结果对GLP-1融合蛋白表达菌株发酵条件的进一步优化提供了依据。  相似文献   

10.
Candida shehatae ATCC 22984, a xylose-fermenting yeast, showed an ability to produce ethanol in both glucose and xylose medium. Maximum ethanol produced by the yeast was 48.8?g/L in xylose and 52.6?g/L in glucose medium with ethanol yields that varied between 0.3 and 0.4?g/g depended on initial sugar concentrations. Xylitol was a coproduct of ethanol production using xylose as substrate, and glycerol was detected in both glucose and xylose media. Kinetic model equations indicated that growth, substrate consumption, and product formation of C. shehatae were governed by substrate limitation and inhibition by ethanol. The model suggested that cell growth was totally inhibited at 40?g/L of ethanol and ethanol production capacity of the yeast was 52?g/L, which were in good agreement with experimental results. The developed model could be used to explain C. shehatae fermentation in glucose and xylose media from 20 to 170?g/L sugar concentrations.  相似文献   

11.
Wu JY  Yeh KL  Lu WB  Lin CL  Chang JS 《Bioresource technology》2008,99(5):1157-1164
Rhamnolipid is one of the most effective and commonly used biosurfactant with wide industrial applications. Systematic strategies were applied to improve rhamnolipid (RL) production with a newly isolated indigenous strain Pseudomonas aeruginosa EM1 originating from an oil-contaminated site located in southern Taiwan. Seven carbon substrates and four nitrogen sources were examined for their effects on RL production. In addition, the effect of carbon to nitrogen (C/N) ratio on RL production was also studied. Single-factor experiments show that the most favorable carbon sources for RL production were glucose and glycerol (both at 40 g/L), giving a RL yield of 7.5 and 4.9 g/L, respectively. Meanwhile, sodium nitrate appeared to be the preferable nitrogen source, resulting in a RL production of 8.6g/L. Using NaNO(3) as the nitrogen source, an optimal C/N ratio of 26 and 52 was obtained for glucose- and glycerol-based culture, respectively. To further optimize the composition of fermentation medium, twenty experiments were designed by response surface methodology (RSM) to explore the favorable concentration of three critical components in the medium (i.e., glucose, glycerol, and NaNO(3)). The RSM analysis gave an optimal concentration of 30.5, 18.1, and 4.9 g/L for glucose, glycerol, and NaNO(3), respectively, predicting a maximum RL yield of 12.6 g/L, which is 47% higher than the best yield (8.6 g/L) obtained from preliminary selection tests and single factor experiments (glucose and NaNO(3) as the carbon and nitrogen source). The NMR and mass spectrometry analysis show that the purified RL product contained L-rhamnosyl-beta-hydroxydecanoyl-beta-hydroxydecanoate (RL1) and L-rhamnosyl L-rhamnosyl-beta-hydroxydecanoyl-beta-hydroxydecanoate (RL2). Meanwhile, HPLC analysis indicates that the molar ratio of RL1 and RL2 in the purified rhamnolipid product was ca. 1:1.  相似文献   

12.
Bowles, Jean A. (University of Colorado School of Medicine, Denver), and William Segal. Kinetics of utilization of organic compounds in the growth of Mycobacterium tuberculosis. J. Bacteriol. 90:157-163. 1965.-To obtain a workable system for a study of the kinetics of nutrient utilization (based on specific quantitative assay) by Mycobacterium tuberculosis, several cultural refinements were introduced: the use of shake culture, a 40-fold increase in the size of inoculum, substitution of glutamate for asparagine as nitrogen source, and elimination of glucose from the medium with glycerol remaining as carbon source. These modifications resulted in reduction to a tenth of the lag phase of glycerol utilization (from 40 to 4 days), and in a greatly increased rate of growth. Both coordinate and sequential patterns of nutrient utilization were in evidence, except in the case of citrate, which was never utilized under a variety of conditions of culture. The coordinate pattern of glucose-glutamate and glucose-glycerol utilization would appear to rule out catabolite repression by glucose. However, elimination of glucose from the medium resulted in elimination of the 4-day lag period before glutamate utilization was initiated, leaving open to question the role of glucose in this system. Evidence is presented for the hypothesis that the sequential pattern of glutamate-glycerol utilization is a function of glutamate repression of glycerol oxidation in the growth of M. tuberculosis, although no diauxie effect is apparent. In a determination of which nutrient-utilization systems were regulated by induction, only in the case of glycerol was evidence obtained for an inducible system. The enzymatic mechanisms underlying these patterns of nutrient utilization are presently being investigated.  相似文献   

13.
以EMP途径与TCA循环中间代谢物的添加为对照,研究在尿素为氮源的产甘油假丝酵母发酵过程中添加氨基酸对甘油产量的影响。结果表明:对甘油产量有强促进作用的氨基酸有谷氨酸、谷氨酰胺、天冬氨酸、天冬酰胺、甘氨酸、赖氨酸、酪氨酸、脯氨酸、组氨酸和丝氨酸,其最适添加浓度在0.26~0.45g/L之间,丙酮酸、α_酮戊二酸、草酰乙酸、柠檬酸和琥珀酸的最适添加浓度在0.24~0.42g/L之间;赖氨酸最适于在0h添加,丙酮酸和草酰乙酸在第14h,谷氨酸、谷氨酰胺、组氨酸、脯氨酸、天冬氨酸、酪氨酸、甘氨酸、α_酮戊二酸和琥珀酸在第30h,天冬酰胺、丝氨酸和柠檬酸在第48h;在最适条件下添加这些促进剂,甘油产量均呈显著增加趋势,转化率和增加率分别达到60%和16%以上。氨基酸的作用机理为其脱氨形成的碳骨架经特定的分解代谢途径进入TCA循环,使其强化,导致碳代谢流在3_磷酸甘油醛节点处发生转移,使甘油合成途径的代谢流增加。  相似文献   

14.
The capability of utilizing a TCA cycle intermediates as the sole carbon source by the multi-vitamin auxotrophic yeastTorulopsis glabrata CCTCC M202019 was demonstrated with plate count method. It is indicated thatT. glabrata could grew on a medium with one of the TCA cycle intermediates as the sole carbon source, but more colonies were observed when glucose, acetate and one of the TCA cycle intermediates coexisted in the medium. Among the intermediates of the TCA cycle examined in this study, cell growth was improved by supplementing oxaloacetate. Further investigation showed that the presence of acetate was necessary when oxaloacetate was supplemented. By supplementing with 10 g/L of oxaloacetate in pyruvate batch fermentation, dry cell weight increased from 11.8 g/L to 13.6 g/L, and pyruvate productivity was enhanced from 0.96 gL−1h−1 to 1.19 gL−1h−1 after cultivation of 56 h. The yield of pyruvate to glucose was also improved from 0.63 g/g to 0.66 g/g. These results indicate that under vitamins limitation, the productivity and yield of pyruvate could be enhancedvia an increase of cell growth by the supplementation of oxaloacetate.  相似文献   

15.
红曲菌(Monascus spp.)是我国重要的药食同源微生物,红曲色素(Monascus pigments,MPs)是其主要次级代谢产物之一。有研究表明,甘油可促进红曲菌产MPs,但作用机制不明。以丛毛红曲菌(Monascus pilosus)MS-1为实验菌株,考察甘油与葡萄糖或蔗糖复合对红曲菌产MPs的影响。在不含碳源的合成培养基中,将甘油与葡萄糖或蔗糖复合,采用分光光度法和高效液相色谱法等分析MPs的产量和组分、生物量及发酵液pH。当甘油与葡萄糖复合,添加甘油后发酵液pH、生物量无显著变化(P0.05),总色价显著降低(P0.05)。当2 g/L或40 g/L甘油与蔗糖复合,发酵液pH显著降低而生物量及总色价显著增加(P0.05)。当40 g/L甘油与蔗糖复合时,总色价是仅以蔗糖为碳源时的16.5倍,且MPs同系物数量明显增多(P0.05)。在合成培养基条件下,甘油促进红曲菌产MPs具有碳源种类的选择性。该结果可为研究甘油影响红曲菌产MPs的作用机制提供参考,为甘油用于MPs生产提供依据。  相似文献   

16.
金黄色破囊壶菌发酵生产DHA的研究   总被引:6,自引:0,他引:6  
顺 4,7,1 0 ,1 3 ,1 6 ,1 9 二十二碳六烯酸 (docosahexaenoicacid ,简称DHA)是ω 3系列多不饱和脂肪酸。近年的研究表明 ,DHA是组成大脑和视网膜的重要结构物质 ,如大脑灰质结构脂质中 6 0 %的脂肪酸均为DHA[1] 。DHA对人体健康有益的生理功能主要表现在[2 ] :调节中枢神经系统功能 ;预防和治疗心血管疾病 ;治疗气喘、关节炎等 ;预防和治疗乳腺癌、结肠癌等。由于DHA具有上述生理功能 ,已在医药、食品、保健品等领域得到广泛应用。目前 ,商品DHA主要来源于深海鱼油 ,如沙丁鱼、金枪鱼等鱼油 ,由于鱼…  相似文献   

17.
Summary Production of the dipeptide antibiotic bacilysin byBacillus subtilis 168 was growth associated and showed no evidence of repression by glucose or sucrose. Carbohydrates other than glucose and sucrose yielded lower specific titers of bacilysin. Bacilysin production in three such carbon sources (maltose, xylose, ribose) was delayed until growth slowed down. Ammonium salts were poor for bacilysin production when used as the sole nitrogen source. When added to the standard medium containing glutamate, they suppressed antibiotic production. Aspartate was slightly better than glutamate for antibiotic production as sole nitrogen source. No other nitrogen source tested, including inorganic, organic or complex, approached the activity of glutamate or aspartate. When added to glutamate, casamino acids, phenylalanine and alanine (a substrate of bacilysin synthetase) suppressed bacilysin production while stimulating growth. Phosphate provided for optimum growth and production at 7.5 mM and both processes were inhibited at higher concentrations. Ferric citrate stimulated growth and inhibited bacilysin production, the effects being due to both the iron and the citrate components. Elimination of ferric citrate stimulated production as did increasing the concentration of Mn to its optimum concentration of 6.6×10–4M.  相似文献   

18.
Lactobacillus rhamnosus is a heterolactic acid bacterium, which can be used to produce flavour compounds like diacetyl and acetoin. Various startegies have been applied to improve the growth rate and diacetyl yield. The use of multiple substrates affected growth as well as the yield of diacetyl. Growth on a medium containing glucose demonstrated a diauxic growth profile, with the second phase of growth being on the product, lactic acid. L. rhamnosus also grew on a medium containing citrate. Growth on medium containing glucose+citrate demonstrated simultaneous utilization of carbon sources. L. rhamnosus did not grow in a medium containing acetate and also did not co-metabolize it with glucose. Maximum specific growth rate ( max) was found to increase in the case of simultaneous utilization of glucose+citrate (0.38 h–1) as compared to glucose as the sole carbon source (0.28 h–1). The yields of diacetyl were also found to increase for glucose + pyruvate and glucose + citrate (0.10 and 0.05 g g–1 of glucose, respectively) as compared to glucose alone (0.01 g g–1 of glucose). The productivity of diacetyl on medium containing glucose and citrate was double that of a medium containing only citrate, although the yields were comparable.  相似文献   

19.
Crude glycerol is a primary by‐product in the biodiesel industry. Microbial fermentation on crude glycerol for producing value‐added products provides opportunities to utilize a large quantity of this by‐product. This study investigates the potential of using the crude glycerol to produce vancomycin (glycopeptide antibiotics) through fermentation of Amycolatopsis orientalis XMU‐VS01. The results show that crude glycerol was the most effective carbon source for mycelium growth and vancomycin production, with 40–60 g/L glycerol concentration as optimal range. Among other culture medium components, potato protein (nitrogen source) and the phosphate concentration had significant effects (p<0.05) for vancomycin production. A Box‐Behnken design and response surface methodology were employed to formulate the optimal medium. Their optimal values were determined as 52.73 g/L of glycerol, 17.36 g/L of potato protein, and 0.1 g/L of dipotassium phosphate. A highest vancomycin yield of 7.61 g/L with biomass concentration of 15.8 g/L was obtained after 120 h flask fermentation. The yield of vancomycin was 3.5 times higher than with basic medium. The results suggest that biodiesel‐derived crude glycerol is a promising feedstock for production of vancomycin from A. orientalis culture.  相似文献   

20.
West TP  Fullenkamp NA 《Microbios》2000,102(402):89-101
The ability of casamino acids and vitamin-assay casamino acids to support gellan production by Sphingomonas paucimobilis ATCC 31461 was examined in a medium containing glucose or corn syrup as the carbon source relative to yeast extract supplementation. When glucose or corn syrup served as the carbon source, the presence of yeast extract in the growth medium stimulated gellan production by strain ATCC 31461 on casamino acids. Using vitamin-assay casamino acids as the nitrogen source, the addition of vitamins lowered gellan synthesis by glucose-grown cells regardless of yeast extract supplementation while gellan elaboration by corn syrup-grown strain ATCC 31461 cells could only be increased by supplementing vitamins into medium lacking yeast extract. Independent of carbon source, the absence of yeast extract in the medium reduced biomass production. Biomass production by the strain grown on either carbon source was increased by supplementing vitamins in the medium containing yeast extract.  相似文献   

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