黑曲霉SL2-111固态发酵生产聚半乳糖醛酸酶

以麸皮为主要原料，采用黑曲霉(Aspergillus niger)诱变菌株SL2—111进行聚半乳糖醛酸酶固态发酵，培养物最高酶活力可达到2695u／g(鲜曲)。产酶最适培养基为：麸皮15g，柚皮粉1．5g，(NH4)2SO4 0．8g，Ca—Cl2 0.075g。最佳产酶条件为：28℃，pH6．0，培养72h。成曲的最佳浸提条件为：以0．1mol／L，pH4．0柠檬酸柠檬酸钠缓冲液为浸提剂，在30℃下浸提5h。     

羊肚菌的液体深层培养

对羊肚菌菌丝体进行液体深层培养 ,以菌丝干重和胞外多糖得率为指标 ,确定了羊肚菌最佳液体种子培养基为 :土豆 1 0 %、蔗糖 3 %、蛋白胨 0 .1 %、酵母膏 0 .5 %、KH2 PO40 0 5 %、MgSO4 0 0 5 % ;羊肚菌发酵的最优培养基为 :蔗糖 4%、麸皮 2 %、NH4 NO30 3 %、KH2 PO4 0 1 5 %、MgSO4 0 1 5 %和酵母膏 0 5 % ;最优培养条件为 :接种量 1 0 %、摇床转速 1 80r/min、温度 2 4℃、pH 5 5。培养 5d ,菌丝产量可达 7 0 0 5g/L ,胞外多糖产量达 1 2 45 g/L。     

沼泽红假单胞菌产类胡萝卜素培养条件的优化研究

对沼泽红假单胞茵产类胡萝卜素的最佳培养基和最佳培养条件进行了研究。结果表明：菌株N，在培养基组成为KH2PO40．6g，K2HPO4 0.9g，酵母膏1．5g，蛋白胨2．0g，NaCl0．5g。自来水1000mL的培养基中，在接种量为3．0％左右，28℃，1500lx，培养基的自然pH值（6．5—8．5）条件下培养5d后，类胡萝卜素产量可达4．84mg／L。     

Sphaerotilus natansFQ40合成聚β-羟基丁酸(PHB)条件的研究

从活性污泥中分离筛选到 1株产PHB的球衣菌FQ40。其最适发酵培养基配方为 (g/L) :蔗糖 1 0 ,牛肉膏 5 ,MgSO4 ·7H2 O 0 2 ,CaCl2 0 0 5 ,FeCl30 0 1 ,K2 HPO4 0 0 4,KH2 PO4 0 0 3 ,NaH2 PO4 ·2H2 O 0 0 5 ,H3BO30 0 0 5。最佳摇瓶发酵条件为 :2 5 0mL三角瓶装 80mL培养液 ,起始pH为 7 0 ,培养温度 3 0℃ ,接种量 1 0 % ,转速 1 5 0r/min ,周期 42h。在优化条件下 ,细胞干重达4 44g/L ,PHB含量为 48% ,PHB浓度为 2 1 3 g/L。     

产胞外多糖泰山羊肚菌液体培养条件的优化

对泰山羊肚菌产胞外多糖 (exopolysaccharides,EPS)的液体培养基组成和发酵条件进行了优化 ,最适碳源是葡萄糖 ,最适氮源是NH4NO3 。正交试验确定最佳培养基组成为麸皮 2 0 0 g/L ,葡萄糖 30 g/L ,NH4NO3 1g/L ,KH2 PO42 g/L ,MgSO4·7H2 O 1 5 g/L。最佳发酵条件为 2 5℃ ,起始 pH值 6 5 ,装液量 10 0mL/瓶 ,接种量10 % ,摇床转速 2 0 0r/min ,发酵时间 4d。在此条件下 ,其胞外多糖含量 (2 135 4 4 1mg/L)比对照(14 5 4 6 39mg/L)提高了 4 6 8%。     

米曲氨基酰化酶液态发酵条件的研究

通过单因素和正交试验 ,对米曲霉 3 0 42产氨基酰化酶摇瓶液态发酵条件进行优化。结果表明 ,较优培养基为 :麸皮 2 %,蛋白胨 1 5 %,玉米浆 0 5 %,酵母膏 0 1 %,KH2 PO4 0 2 %,无水MgSO4 0 1 %,吐温 80 0 0 5 %。酶活最高达到 1 2 5 8u/g(湿菌体 ) ,通过对培养条件的研究 ,最佳起始 pH 7.0、温度 2 6℃、装液量 1 0 0mL/L三角瓶。     

真菌α-淀粉酶产生菌的筛选及其固态发酵条件的初步研究

从酒曲和酒药中分离筛选出一株产α-淀粉酶的菌株A019。经形态学初步鉴定为米曲霉。研究了碳源、氮源、培养温度与时间等因素对A019产酶的影响,并用响应面法对培养基组成进行了优化。最佳的培养基组成和培养条件为:麸皮8g,豆饼粉2.28g,酵母膏0.1g,K2HPO4·3H2O1%,MgSO4·7H2O0.094%,加水量12.9ml,培养基的起始pH5.0左右,28℃培养48h。在此条件下α-淀粉酶的酶活达到410U/g干曲左右。     

浓香乳菇深层发酵工艺的研究

报道了浓香乳菇液体发酵的试验结果。研究了 7种营养物质对菌丝生长的影响 ,应用正交试验得到了发酵培养基 ,确定了摇瓶发酵条件 ,进行了 3L发酵罐培养的初步试验。结果表明 ,其适宜发酵培养基的组成为麸皮汁 4%、土豆汁 30 %、玉米粉 5 %、KH2 PO40 3%、MgSO40 1 5 %、VB1 0 0 5 % ;适宜的发酵条件是 2 4℃、培养基起始 pH 4 0、摇瓶装量 80mL/30 0mL三角瓶、接种量 1 0 %、大号玻璃珠 1 0个 /瓶 ;产菌丝干重 1 95 g/1 0 0mL左右。     

青霉菌Y3固态发酵产酶条件的研究

筛选了一株高产木聚糖酶及纤维素酶的青霉菌 (Penicilliumemersonii)Y3 菌株 ,研究了其在固体培养基中的发酵条件。该菌株的最佳培养条件为 :起始pH值为 4 .6 ,发酵温度为 2 9℃ ,每 1g固体培养基中接种量为 1ml孢子悬液 (孢子悬液的浓度为 2 .5× 10 7个 (孢子 ) /ml) ,玉米芯与麦麸比为 7∶3,尿素为氮源 ,发酵 3d。木聚糖酶活力可达 4 2 36U/g ,纤维素酶活力可达 1895U/g。粗酶的最适反应温度为 5 0℃ ,pH值为 5 .0 ,热稳定性在 5 0℃以下。     

基因工程菌Pichia Pastoris高密度培养条件的摇瓶研究

对基因工程茵Pichia pastoris高密度培养的培养基进行了研究，选取了价廉易得的培养基，对此培养基各组分进行了优化，并对其它发酵条件进行了优化试验，得到了最优培养基配方及培养条件。结果表明，培养基最佳配方为：葡萄糖为5％，氨水单次补料量为20μL(250mL摇瓶装液量为20mL)，KH2PO4浓度为0．7％，培养基其它组分为：K2HPO40．1％、MgSO4．7H2O0．03％、FeSO4．7H2O0．05％、MnSO4．H2O0．05％。种子液最佳培养时间为40h，接种量为10％，250mL三角瓶装液量为20mL，摇床转速为220r／m，发酵培养基最佳初始pH5．5，发酵温度为30℃，发酵结束时间64h。在此优化的培养基及培养条件下，茵体密度达到最高，OD600达到64．3，细胞干重20．2g／L。     

木聚糖酶产生菌的筛选

目的 从土壤中筛选木聚糖酶产量高的菌株,并研究其产酶的初步条件.方法 根据培养基上水解圈的大小和摇瓶培养后木聚糖酶的酶活性,筛选产酶量高的菌株.通过一系列单因素试验,研究影响产酶的碳源、氮源和无机盐.结果 筛选到1株产木聚糖酶的菌株,其产酶的较优碳源、氮源和无机盐为麸皮(30 g/L)、牛肉膏(10g/L)和磷酸氢二钾(1.0 g/L).结论 筛选到的菌株具有生产木聚糖酶的潜力.     

嗜热拟青霉利用玉米芯产木聚糖酶的发酵条件优化

以一株新的嗜热拟青霉(Paecilomyces thermophila)J18为出发菌株,对其液态发酵产木聚糖酶的条件进行了优化。结果表明,以玉米芯为碳源能高效诱导木聚糖酶的胞外分泌,胰蛋白胨为最佳氮源,初始pH和培养温度分别为pH 7.5,50℃,所得酶活力最高。在优化后的培养基和培养条件下,第5 d酶活力达到峰值,为1 276 U/mL,表明该拟青霉能够利用农业废弃物高效生产木聚糖酶。该酶水解桦木木聚糖主要生成木二糖和木三糖,适合生产低聚木糖。     

Bacillus pumilus WL-11蛋白酶对其木聚糖酶合成的影响

研究了不同的营养条件下短小芽孢杆菌WL-11产蛋白酶和木聚糖酶的情况。结果表明,碳源对WL-11木聚糖酶的合成具有诱导或抑制作用,对其蛋白酶的合成影响并不显著;氮源对WL-11木聚糖酶的合成不具有直接的调节作用,而可能通过调节其蛋白酶的合成量来间接影响其木聚糖酶的合成,且两者的合成量之间存在一定的正相关。     

木聚糖酶产生菌的筛选

目的从土壤中筛选木聚糖酶产量高的菌株,并研究其产酶的初步条件。方法根据培养基上水解圈的大小和摇瓶培养后木聚糖酶的酶活性,筛选产酶量高的菌株。通过一系列单因素试验,研究影响产酶的碳源、氮源和无机盐。结果筛选到1株产木聚糖酶的菌株,其产酶的较优碳源、氮源和无机盐为麸皮(30 g/L)、牛肉膏(10 g/L)和磷酸氢二钾(1.0 g/L)。结论筛选到的菌株具有生产木聚糖酶的潜力。     

木聚糖酶的研究进展

木聚糖是仅次于纤维素的第二丰富的可再生资源,通过筛选出产木聚糖酶的菌株,利用基因工程技术,将木聚糖酶基因进行异源表达,提高木聚糖酶产量。该文综述了菌株产生的木聚糖酶酶学性质,并对酶学性质进行改善使木聚糖酶更符合应用的条件,同时研究木聚糖酶分离纯化的步骤,来提高木聚糖酶的酶活。文章重点介绍了木聚糖酶的产生菌和木聚糖酶在基因工程技术等方面研究进展,并对木聚糖酶的分离纯化方法做了简要地介绍。     

Comparison of three expression systems for heterologous xylanase production by S. cerevisiae in defined medium

The influence of the auxotrophic deficiencies of the host strain and expression vector selection on the production of a heterologous protein was investigated. Heterologous xylanase production by two prototrophic S. cerevisiae transformants, containing either a plasmid-based, YEp-type expression system or an integrative, YIp-type expression system, were compared with production by an auxotrophic transformant, containing an identical YEp-type expression system, in batch and continuous cultivation, using a chemically defined medium. Heterologous xylanase production by the auxotrophic strains in defined medium was critically dependent on the availability of amino acids, as extracellular xylanase production increased dramatically when amino acids were over-consumed from the medium to the point of saturating the cell. Saturation with amino acids, indicated by an increased leakage of amino acids from the cell, was thus a prerequisite for high level of heterologous protein production by the auxotrophic strain. Maximal xylanase production levels by the auxotrophic strain corresponded to the levels obtained with a similar prototrophic strain during cultivation in defined medium without amino acids. Superfluous auxotrophic markers thus had a strong deleterious effect on heterologous protein production by recombinant yeasts, and the use of such strains should be limited to initial exploratory investigations. The increased copy number and foreign gene dosage of the YEp-based expression vector, stabilized by the ura3 fur1 autoselection system, significantly improved production levels of heterologous xylanase, compared to the YIp system, which is based on a single integration into the yeast genome. No evidence was found of the possible saturation of the host secretory capacity by multicopy overexpression. Stable production of heterologous xylanase at high levels by the prototrophic YEp-based recombinant strain, compared to the YIp system, was demonstrated.     

短小芽孢杆菌固态发酵生产木聚糖酶的培养条件优化

采用单因素试验和正交试验的方法研究了短小芽孢杆菌固态发酵生产木聚糖酶的培养条件,确定了利用麸皮作为在主要基质进行固态发酵生产木聚糖酶的适宜条件：pH9、温度35℃、料液比1：2、接种量10％。在500mL三角瓶中固态培养72h左右,木聚糖酶的活力可达4915U／g干曲。     

Production of Xylanase with a transformant of Aspergillus oryzae RIB40 in a Liquid-Surface Immobilization (LSI) System

Xylanase production by a XynF1 (33 kDa)-transformant of Aspergillus oryzae RIB40 was compared between submerged cultivation (SmC) and liquid-surface immobilization (LSI) systems. While the accumulation of xylanase in the SmC decreased by prolonged incubation, LSI system enabled the continuation of xylanase production to afford 4.5-fold xylanase production compared with the SmC system.     

Production,Characterization and Application of a Xylanase from Streptomyces sp. AOA40 in Fruit Juice and Bakery Industries

Streptomyces sp. AOA40, which produces halotolerant and thermotolerant xylanase, was isolated from Mersin soil. Various carbon sources were tested for xylanase production with selected fermentation medium. The best carbon source was selected as corn stover. The effect of corn stover concentration and particle size, composition of fermentation medium, fermentation condition such as initial pH and agitation rate on xylanase production was determined. After production, xylanase was partially purified with ion-exchange chromatography and gel filtration chromatography for characterization of xylanase and application in fruit juice and dough improvement. The optimum pH for the activity of xylanase occurred at pH 6.0 in phosphate buffer, while the optimum temperature was 60°C. The relative xylanase activity in the pH ranges of 4–9 remained between 59.93 and 54.43% of the activity at pH 6.0 (100.00%). The xylanase activity showed a half-life of 172 min at 70°C, which was reduced to 75 min at 80°C. The enzyme was highly inhibited by 10–100 mM of Hg⁺², EDTA, Mg⁺², SDS and 100 mM Cu⁺². Clarity of fruit juices increased after enzymatic treatment of apple (17.85%), grape (17.19%) and orange juice (18.36%) with partially purified xylanase and also reducing sugar concentrations of these fruit juices were improved by 17.21, 16.79 and 19.57%, respectively. Also, dough volume was raised 17.06% with using partially purified Streptomyces sp. AOA40 xylanase in bread making.     

嗜热棉毛菌固体发酵产木聚糖酶条件的优化

通过单因素实验优化了嗜热棉毛菌CAU44利用农业废弃物固体发酵产木聚糖酶的发酵条件。结果表明,发酵产酶的最佳碳源为玉米芯和麦麸以4∶6混合的复合碳源,最佳氮源为(NH4)2SO4,最佳水分含量为80%,培养基最佳初始pH为4.0,最佳表面活性剂及添加量为0.5%的Tween-60。在优化后的发酵条件下50℃培养7d,嗜热棉毛菌CAU44产木聚糖酶的酶活力最高,达到20343U/g干基碳源,为目前国内报道的最高值。因此,嗜热棉毛菌CAU44在利用农业废弃物固体发酵产木聚糖酶方面有很大的应用前景。