不同碳源对须糖多孢菌生长发育及丁烯基多杀菌素生物合成的影响

本文研究了不同碳源对须糖多孢菌生长以及丁烯基多杀菌素生物合成的影响,通过寻找优势碳源优化发酵培养基配方,促进须糖多孢菌丁烯基多杀菌素的生物合成。试验共设11个处理,1个对照,通过单因素试验比较不同处理组菌体OD600值和丁烯基多杀菌素产量,筛选获得最优碳源及其发酵培养基配方。结果表明,除可溶性淀粉和木糖外,须糖多孢菌在9种碳源中都能进行生长,对不同构型碳源显示较好的利用率。在以半乳糖、葡萄糖、果糖和甘露糖作为碳源时具有较好的生长速率,而以甘露糖为碳源时能显著促进丁烯基多杀菌素的合成。选择甘露糖最佳添加浓度为5 g/L,须糖多孢菌最高菌体浓度和丁烯基多杀菌素产量分别是初始配方条件的1. 32倍和1. 78倍,显著提高了丁烯基多杀菌素的产量。上述结果为培养基碳源对丁烯基多杀菌素生物合成影响机制的研究及丁烯基多杀菌素大规模工业化发酵生产提供了科学依据和新的技术途径。     

斯达氏油脂酵母利用混合糖发酵产油脂

研究了斯达氏油脂酵母Lipomyces starkeyi2#利用葡萄糖-木糖混合糖为碳源生长和油脂积累特性。L.star-keyi2#利用70 g/L葡萄糖和70 g/L木糖作为碳源在30℃下摇瓶发酵96 h,糖利用率均达90%以上,菌体生物量分别为14.1 g/L和13.1 g/L,油脂质量分数分别为55.7%和52.6%。相同条件下该菌株利用混合糖(葡萄糖46 g/L,木糖24 g/L)为碳源时总糖利用率、生物量和油脂质量分数分别为75.1%,15.0 g/L和40.0%。借助于P lackett-Burm an设计法和单因子实验法对培养条件进行了优化,结果表明发酵96 h混合糖利用率可达到97.3%,发酵120 h后混合糖利用率、生物量和菌体油脂质量分数分别达99.5%、19.0 g/L和52.6%。生物量得率和油脂得率分别达到27%和14%。     

Candida glycerinogenes不同碳源代谢的初步研究

研究了不同碳源对Candidaglycerinogenes的菌体生长、发酵液pH值及代谢产物的影响,结果发现以葡萄糖、果糖等单糖为碳源时茵体生长较快,最终生物量比以蔗糖、麦芽糖等二糖为碳源时高20％～30％;导致发酵前12h发酵液pH值明显下降的主要因素是乳酸;与葡萄糖为碳源转化为甘油相比,果糖为碳源时更易累积乙醇;以蔗糖、麦芽糖为碳源时,用于转化生成甘油的碳源明显降低,碳源主要用于茵体自身生物合成及HMP途径,以蔗糖为碳源时,用于乳酸、丙酸及柠檬酸生物合成的碳源较麦芽糖明显提高,TCA途径代谢较为活跃。     

Candida glycerinogenes metabolic fermentation using different carbon sources

研究了不同碳源对Candida glycerinogenes的菌体生长、发酵液pH值及代谢产物的影响,结果发现以葡萄糖、果糖等单糖为碳源时菌体生长较快,最终生物量比以蔗糖、麦芽糖等二糖为碳源时高20％～30％;导致发酵前12 h发酵液pH值明显下降的主要因素是乳酸;与葡萄糖为碳源转化为甘油相比,果糖为碳源时更易累积乙醇;以蔗糖、麦芽糖为碳源时,用于转化生成甘油的碳源明显降低,碳源主要用于菌体自身生物合成及HMP途径,以蔗糖为碳源时,用于乳酸、丙酸及柠檬酸生物合成的碳源较麦芽糖明显提高,TCA途径代谢较为活跃.     

麦白霉素发酵培养基的改进

研究了淀粉水解液全部替代葡萄糖作为碳源发酵麦白霉素的工艺条件。用正交实验得出摇瓶培养基消毒后的最佳基质浓度为总糖5.0g/100mL,还原糖3.8g/100mL,氨基氮60mg/mL,溶解磷300μg/mL。以50L发酵罐进行实验,得出了麦白霉素的发酵代谢曲线和淀粉的适宜水解条件。经30t发酵罐放大实验结果表明,与葡萄糖作碳源相比,用淀粉水解液作碳源发酵麦白霉素,发酵单位提高23.0％,生产成本降低38.0％。     

嗜热子囊菌利用短链有机酸生产角质酶

以嗜热子囊菌(Thermobifida fusca WSH03-11)发酵生产角质酶为模型，研究微生物利用市政污泥厌氧酸化所产短链有机酸为碳源发酵生产高附加值产品的可能。发现：(1)以丁酸、丙酸和乙酸为碳源时，有机酸和氮元素浓度分别为8.0 g/L和1.5 g/L有利于角质酶的生产；而以乳酸为碳源时，最适有机酸和氮源浓度分别为3.0 g/L和1.0 g/L；(2)改变诱导物角质的浓度，以丁酸、丙酸、乙酸和乳酸为碳源，分别比优化前提高了31.0%、13.3%、43.8%和73.2%；(3)在四种有机酸中，T. fusca WSH03-11利用乙酸的速率最快，平均比消耗速率是丙酸的1.3倍，丁酸的2.0倍及乳酸的2.2倍；以丁酸为碳源时的酶活(52.4 U/mL)是乳酸的1.7倍、乙酸的2.5倍和丙酸的3.2倍；角质酶对乳酸的得率(12.70 u/mg)分别是丁酸的1.4倍、丙酸的3.0倍和乙酸的3.8倍；(4)以混合酸为碳源生产角质酶，T. fusca WSH03-11优先利用乙酸，而对丁酸的利用受到抑制。进一步研究发现，混合酸中0.5 g/L的乙酸将导致丁酸的消耗量降低66.7%。这是首次利用混合酸作碳源发酵生产角质酶的研究报道。这一研究结果进一步确证了利用市政污泥厌氧酸化所产有机酸为碳源发酵生产高附加值产品的可行性，为以廉价碳源生产角质酶奠定了良好的基础。     

虎掌菌菌丝富硒培养的研究

为生产合适的硒源提供一种思路,以菌丝体生物量、含硒量、还原糖、氨态氮和蛋白质为指标,采用四因素三水平正交设计法优化虎掌菌的富硒发酵条件,探讨不同浓度的硒对虎掌菌固体培养菌丝生长和液体培养产生的生物组分的影响。结果表明,高浓度的硒抑制虎掌菌菌丝体生长;正交试验选择不同的衡量指标,由极差分析得出各因素的影响程度大小,结合证实试验得到以还原糖为指标的最优组合为葡萄糖6%(质量分数)、酵母浸膏3%(质量分数)、KH_2PO_4 0.1%(质量分数)、Na_2SeO_3 0.6 mmol/L,其菌丝体生物量和氨态氮含量较高。与对照相比,加硒后虎掌菌发酵液中氨态氮、还原糖和总糖含量显著增加(P0.05);当硒浓度为0.5 mmol/L时,氨态氮、还原糖和总糖含量均达到最高值。菌丝体生物量和可溶性蛋白质分别在硒浓度为0.2 mmol/L和0.4 mmol/L时达到最大值。虎掌菌富硒培养后,发酵液的营养成分含量会发生变化。     

嗜热子囊菌利用短链有机酸生产角质酶

以嗜热子囊菌(Thermobifida fusca WSH03-11)发酵生产角质酶为模型，研究微生物利用市政污泥厌氧酸化所产短链有机酸为碳源发酵生产高附加值产品的可能。发现：(1)以丁酸、丙酸和乙酸为碳源时，有机酸和氮元素浓度分别为8.0 g/L和1.5 g/L有利于角质酶的生产；而以乳酸为碳源时，最适有机酸和氮源浓度分别为3.0 g/L和1.0 g/L；(2)改变诱导物角质的浓度，以丁酸、丙酸、乙酸和乳酸为碳源，分别比优化前提高了31.0%、13.3%、43.8%和73.2%；(3)在四种有机酸中，T. fusca WSH03-11利用乙酸的速率最快，平均比消耗速率是丙酸的1.3倍，丁酸的2.0倍及乳酸的2.2倍；以丁酸为碳源时的酶活(52.4 U/mL)是乳酸的1.7倍、乙酸的2.5倍和丙酸的3.2倍；角质酶对乳酸的得率(12.70 u/mg)分别是丁酸的1.4倍、丙酸的3.0倍和乙酸的3.8倍；(4)以混合酸为碳源生产角质酶，T. fusca WSH03-11优先利用乙酸，而对丁酸的利用受到抑制。进一步研究发现，混合酸中0.5 g/L的乙酸将导致丁酸的消耗量降低66.7%。这是首次利用混合酸作碳源发酵生产角质酶的研究报道。这一研究结果进一步确证了利用市政污泥厌氧酸化所产有机酸为碳源发酵生产高附加值产品的可行性，为以廉价碳源生产角质酶奠定了良好的基础。     

Alkalitalea saponilacus产木聚糖酶的碳源优化及酶学特性

【目的】确定厌氧盐碱细菌Alkalitalea saponilacus产木聚糖酶所需的碳源,优化木聚糖粗酶的提取条件并分析酶学性质。【方法】应用GC技术分析A.saponilacus发酵木聚糖的主要产物;利用二硝基水杨酸法(DNS)测定木聚糖酶活力以获得最优的碳源、提取粗酶的最佳条件及其酶学特性。【结果】A.saponilacus以不同来源木聚糖为底物时,发酵产生的主要产物丙酸含量都在80%以上。若以0.4%(W/V)蔗糖+0.1%(W/V)桦木木聚糖为复合碳源时,木聚糖酶活力是以桦木木聚糖或者蔗糖为单一碳源时的3.2倍。木聚糖酶的酶活力在盐度2%–6%、pH 7.0和55°C达到最佳且在该条件下的酶活力为590 IU/mg。此外,该酶活力在0.2%Tween 20存在时增加,而在5 mmol/L Mg~(2+)和0.2%Triton X-100存在时无显著影响,但在Cu~(2+)、Fe3+和Ni~(2+)等金属离子存在时则被显著抑制。【结论】A.saponilacus发酵主产物丙酸以及生物合成的木聚糖酶在工业生产中具有广泛的应用前景。     

鸡菌液体深层发酵的研究

本文研究了不同碳源、氮源和起始pH对鸡菌液体深层发酵的影响。采用均匀设计方法对发酵培养基进行优化,发现当培养基的组成为大米水解糖4.29%, KH2PO4 0.31%,酵母粉0.53%,pH 4.5时,在28℃、150r/min的条件下培养5d,生物量可高达19.8g/L。     

Growth and fermentation patterns of Saccharomyces cerevisiae under different ammonium concentrations and its implications in winemaking industry

AIMS: To study the effects of assimilable nitrogen concentration on growth profile and on fermentation kinetics of Saccharomyces cerevisiae. METHODS AND RESULTS: Saccharomyces cerevisiae was grown in batch in a defined medium with glucose (200 g l(-1)) as the only carbon and energy source, and nitrogen supplied as ammonium sulphate or phosphate forms under different concentrations. The initial nitrogen concentration in the media had no effect on specific growth rates of the yeast strain PYCC 4072. However, fermentation rate and the time required for completion of the alcoholic fermentation were strongly dependent on nitrogen availability. At the stationary phase, the addition of ammonium was effective in increasing cell population, fermentation rate and ethanol. CONCLUSIONS: The yeast strain required a minimum of 267 mg N l(-1) to attain complete dryness of media, within the time considered for the experiments. Lower levels were enough to support growth, although leading to sluggish or stuck fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings reported here contribute to elucidate the role of nitrogen on growth and fermentation performance of wine yeast. This information might be useful to the wine industry where excessive addition of nitrogen to prevent sluggish or stuck fermentation might have a negative impact on wine stability and quality.     

Influence of carbon and nitrogen sources on antifungal metabolite production by bacterium associated with entomopathogenic nematode against Penicillium expansum

A specific symbiotic Bacillus sp. isolated from a rhabditid entomopathogenic nematode, Rhabditis (Oscheius) sp. was found to produce large number of bioactive compounds. The present study was conducted to determine the effect of carbon and nitrogen sources for the production of antimicrobial substances by Bacillus sp. The yield of the crude antimicrobial substances and antimicrobial activity against the test micro-organism also differed significantly when carbon and nitrogen sources in the fermentation media were changed. The antifungal activity was significantly high in yeast extract plus fructose (46.5?±?2.12?mm) followed by yeast extract plus maltose, beef extract plus fructose and meat infusion plus glucose. High pressure liquid chromatography analysis of the crude antimicrobial substances revealed different peaks with different retention time indicating that they produced different compounds. When the carbon source was not included in the fermentation media, the antimicrobial production was substantially reduced. The results indicate that carbon source in the fermentation media plays a vital role in the production of antifungal substances. It is concluded that yeast extract and fructose as nitrogen and carbon sources produced maximum activity, which can effectively control the blue mould caused by Penicillium expansum in apples and pears.     

Utilization of Carbon- and Nitrogen-containing Compounds for Neomycin Production by Streptomyces fradiae

A number of carbon and nitrogen compounds were tested for their effect on growth of Streptomyces fradiae 3535 and neomycin production. Dextrin, starch, and maltose were excellent carbon sources for neomycin production, and sodium nitrate, aspartic acid, and glutamic acid were adequate nitrogen sources. Studies on biochemical changes during fermentation in four typical media indicated that there was no direct relation between the growth of the organism and neomycin formation. The pH of the medium might be an important factor for neomycin synthesis. The quantitative formation of neomycin components depended on the variation of carbon and nitrogen sources. On the basis of this study, a suitable synthetic medium for neomycin production has been developed.     

Xylanase production by Ganoderma lucidum on liquid and solid state fermentation

Ganoderma lucidum, a white rot fungus, was exploited for its potentials to produce xylanase employing shake and solid-state culture conditions. Different culture conditions such as pH, temperature, carbon and nitrogen requirements for its growth and production of xylanase were optimized. The culture media pH 6.0-7.0 and temperatures 30 degrees-35 degrees C significantly promoted the growth as well as xylanase secretion into the media. Xylan and peptone were found to be the suitable carbon and nitrogen sources. Among the different agrowastes used, wheat bran was found to be the best substrate for the test fungus for the production of xylanase than sugarcane bagasse and rice bran in solid-state fermentation.     

娄彻氏链霉菌ATCC10739产抗生素Borrelidin发酵条件优化及其分离纯化

对娄彻氏链霉菌ATCC10739固体发酵产生十八元大环内酯抗生素Borrelidin进行了发酵条件的优化。首先筛选得到了理想的发酵培养基;其次考察了发酵时间、起始pH值以及在ISP-2培养基中添加附加碳源、氮源对Borrelidin产量的影响。初步确定最适发酵条件为:ISP-2培养基中添加1%甘油,起始pH值为6.0,培养温度30°C,发酵时间为7 d,产量可达1.336 mg/L。采用有机溶剂萃取、硅胶层析和半制备型高效液相色谱(HPLC)等分离技术纯化得到Borrelidin。     

Fermentative production of spiramycins

Fermentative production of spiramycins by Streptomyces ambofaciens has been performed using fermentation media of different chemical compositions. Medium I was selected from nine media as the best for production of high titres of spiramycins. Biochemical changes which occurred during fermentative production of spiramycins revealed that adjustment of the initial pH value of the medium was very important. The initial pH value of the fermentation medium which allowed the organism to produce a good yield of antibiotic was 6.5. The fermentation period affected the formation of spiramycins, and the maximum incubation period required for the fermentation process was 120 h. The role of inoculum on spiramycin yield showed that it was better to inoculate the fermentation medium with vegetative cells of Streptomyces ambofaciens rather with spores. The carbon source influenced spiramycin biosynthesis: dextrin was the best carbon source and stimulated the organism to form high titres of antibiotics. The best concentrations of dextrin and glucose for increased antibiotic yields were 25 and 15 gl^?1, respectively. Organic sources in the fermentation medium were more efficient than inorganic nitrogen sources for spiramycin formation. Fodder yeast was the best organic nitrogen source in fermentative production of spiramycins. The maximal concentrations of fodder yeast, soybean meal, peptone, Ca(NO₃)₂ and NH₄NO₃ for increased antibiotic yield were 6.5, 6.0, 4.0, 10.0 and 4.0 gl^?1, respectively.     

Impact of beef extract and six carbon source on antifungal metabolites production by bacterium associated with entomopathogenic nematode against Fusarium oxysporum

A specific symbiotic Bacillus species isolated from a rhabditid entomopathogenic nematode, Rhabditis (Oscheius) sp., was found to produce a number of bioactive compounds. The present study was conducted to determine the effect of six different carbon sources in combination with beef extract on the production of antifungal substances by Bacillus sp. The yield of crude antimicrobial substances and antimicrobial activity against the test microorganism also differed significantly when the carbon sources in the fermentation media were changed. The highest yield was recorded for fructose plus beef extract (956?mg/l). The antifungal activity was significantly high in beef extract plus maltose (21?±?1.5?mm) followed by beef extract plus glucose and beef extract plus fructose. Antifungal activity was significantly reduced in beef extract plus lactose and sucrose. High pressure liquid chromatography analysis of the crude antimicrobial substances revealed different peaks with different retention times indicating that they produced different compounds. When a carbon source was not included in the fermentation media, the antifungal production was substantially reduced. Carbon source in the fermentation medium plays a vital role in the production of antimicrobial substances. Beef extract and maltose as nitrogen and carbon sources in the fermentation medium produced maximum antifungal activity. It is concluded that Beef extract and maltose as nitrogen and carbon sources produced maximum activity which can effectively control the Fusarium oxysporum which causes vascular fusarium wilt in tomato, tobacco, legumes, cucurbits, sweet potatoes, banana, etc.     

The fermentative production of acetone-butanol by Clostridium acetobutylicum.

Fourteen different media were used in the fermentative production of acetone-butanol. The highest total yields were achieved in medium I. Potato starch and soluble starch were suitable as carbon sources. The best concentrations of potato starch and soluble starch were 500.0 and 10.0 g/l, respectively. Peptone was the most favourable nitrogen source. The best concentration of peptone was 4.0 g/l. Calcium carbonate in 3.6 g/l acted as buffering agent in the fermentation process. The best initial pH value of the fermentation medium was 6.0. The optimum temperature was 32--33degreesC. The fermentation process required 120 h to obtain maximum yields of acetone-butanol.     

费氏链霉菌中子辐射诱变菌株对新霉素的生物转化

对新霉素产生菌费氏链霉菌进行中子辐射诱变和突变株筛选, 获得不产新霉素的突变株。以突变株为转化菌种, 以新霉素为底物, 对转化发酵液进行高效液相色谱分析, 研究了不同转化条件对新霉素转化的影响。结果表明, 底物浓度、底物添加时间、底物添加方式、接种量、培养基装量、转化时间、碳源、氮源、pH、温度对新霉素转化具有不同程度的影响。以转化条件优化参数进行转化大量培养, 转化液经4步离子交换层析进行分离纯化, 薄层层析检测纯化样品为单一斑点。采用薄层生物自显影对获得的4个转化产物分离样品做生物活性检测, 发现4个样品对金黄色葡萄球菌和姜青枯假单孢杆菌都具有抑制活性, 1个样品对大白菜软腐样品具有明显的抑制活性。     

Organic and inorganic nitrogen source effects on the metabolism of Clostridium acetobytulicum

Summary The effects of organic and inorganic nitrogen combinations on cell growth, solvent production and nitrogen utilization by Clostridium acetobutylicum ATCC 824 was studied in batch fermentations. Fermentations in media with 10 mM glutamic acid, as the organic nitrogen source, and 0 mM to 10 mM ammonium chloride, as the inorganic nitrogen source had a solvent yield of 0.8 to 1.08 mmol solvent/mmol glucose used, with a slow fermentation rate (2 mmol solvent/l h^-1). When media contained 20 mM or 30 mM glutamic acid as well as 2.5 to 7.5 mM ammonium chloride the fermentation rate increased (5.5 mmol/l h^-1) while the solvent yield remained constant (0.86 to 0.96 mmol solvent/mmol glucose used). Total solvent production was higher in media containing 20 mM or 30 mM glutamic acid than with 10 mM glutamic acid.