桃儿七内生真菌产鬼臼毒素发酵条件研究

对鬼臼毒素(Podophyllotoxin)产生菌Ty的发酵培养基及发酵条件进行优化研究。结果表明。优化的半合成培养基组成为:可溶性淀粉2%,蔗糖2%.蛋白胨0.1%,酵母膏0.1%,K_2HPO_4 0.1%.NaCl 0.1%,pH 6.5,装液量80 mL/250 mL三角瓶,接种量2.5%,发酵培养6d,细胞干重为8.5g/L,鬼臼毒素产量达2.418μg/L.     

法夫酵母产虾青素培养基及培养条件的优化

采用单因素实验对高产虾青素的法夫酵母(Phaffia rhodozymaJMU-MVP14)的培养基及培养条件进行优化,确定最佳培养基组成是:葡萄糖30g/L,酵母膏6g/L,(NH4)2SO41.015g/L(C/N=65),MgSO4·7H2O1g/L,KH2PO413.6g/L(C/P=10),CaCl20.2g/L。最佳培养条件是:初始pH7.0,接种量5%,装液量30mL,培养温度20℃。在此条件下,于7L发酵罐、进行发酵培养,其生物量和虾青素含量分别为13.13g/L和67.07mg/L,与初始培养条件相比分别提高了85.8%和15.8%。     

大肠杆菌产β-内酰胺酶培养基条件

对大肠杆菌(Escherichia coli)产β-内酰胺酶培养基条件进行优化研究,以提高β-内酰胺酶活力。研究了碳源、混合碳源配比、氮源、混合氮源配比、氯化钠、微量元素等因素对菌体干重及β-内酰胺酶活力的影响。确定菌种最佳培养时间为40h,最佳培养基组成为:葡萄糖16g/L、可溶性淀粉4g/L、蛋白胨20g/L、酵母浸粉10g/L、氯化钠6g/L、Zn2+浓度0.9 mg/L,β-内酰胺酶活力达2 648.95 nkat,比培养基优化前提高了18%。     

产谷胱甘肽重组巴斯德毕赤氏酵母发酵条件的研究

考察了培养基组成及培养条件对重组巴斯德毕赤氏酵母(Pichia pastoris)x-33(pGAPZA-gsh 1)合成谷胱甘肽(GSH)的影响。优化后的培养基组成为:甘油30g/L、蛋白胨40g/L、酵母膏9g/L、半胱氨酸0.36 g/L、KH_2PO_4 3g/L;培养条件为:自然pH、摇床转速200r/min、装液量为30mL/250mL、接种量为10%。在此优化条件下重组酵母的GSH产量是98.5mg/L,为优化前的2.33倍,生物量最大值达到19.6g/L。在5L发酵罐上进行放大实验,发酵结束后GSH产量、重组菌的生物量分别为97.9mg/L,18.7g/L与摇瓶发酵结果基本吻合。     

地衣芽孢杆菌产Levan果聚糖发酵条件的优化

通过单因素试验(培养基用水、碳源、氮源、培养温度和培养基初始pH值)和正交试验对地衣芽孢杆菌(Bacillus licheniformis)8-37-0-1发酵产生Levan果聚糖的培养基组成及培养条件进行优化,采用苯酚-硫酸法测定多糖含量。结果表明：以蔗糖100g/L、牛肉膏1.0g/L、酵母粉0.6g/L、K2HPO4 3.0g/L、KH2PO4 3.0g/L、NaCl 1.0g/L、MgSO4·7H2O 0.2g/L、FeSO4·7H2O 0.001g/L,自来水配制,培养基初始pH5.0,30℃培养8-37-0-1菌株24h,Levan果聚糖产量达到最高值41.7g/L,约是未优化时的5.0倍。     

产γ-亚麻酸诱变菌拉曼被孢霉HLY0902的发酵条件优化

采用正交实验和单因素实验,分别对诱变菌拉曼被孢霉HLY0902的培养基及培养条件进行优化,以期提高γ-亚麻酸(GLA)的产量。结果表明:当发酵培养基组成为葡萄糖100 g/L、酵母浸粉10 g/L、KH_2PO_44 g/L、Na NO_31 g/L、Mg SO_4·7H_2O 0.5 g/L时,GLA的产量最大,可达1.05g/L,较优化前提高了43.8%;最优培养条件为接种量10%,装液量20%,发酵时间168 h,适合菌体生长和油脂积累的p H为5.5、发酵温度为22℃,适合GLA积累的p H为7.5、发酵温度为20℃。通过该系列的优化研究,诱变菌拉曼被孢霉HLY0902产GLA的能力显著提高。     

产谷胱甘肽荧光假单胞菌发酵条件的优化

方法:在实验室前期工作中,分离得到一株可生产谷胱甘肽(GSH)的荧光假单胞菌BJYG12,在单因素的基础上,通过正交试验对该菌株发酵生产GSH的条件进行了优化。结果:发酵温度28℃,初始pH 7.5,接种量5%,培养基含酵母膏25g/L,蔗糖15g/L,KH_2PO_4 1.5g/L,MgSO_4 2.5g/L,优化发酵条件后,菌株产GSH的能力增长了15.7%。     

刺孢小克银汉霉发酵产γ-亚麻酸时补料工艺研究

研究刺孢小克银汉霉(Cunninghamellaechinulata)发酵生产γ-亚麻酸(GLA)的补料工艺。结果表明,该菌株在含有0.25%黄豆饼粉的发酵培养基中,以分批补料方式,即培养2.5d后补入20g/L食用糖与10g/L麦芽糖混合液,培养第3d时补入1.0g/L(NH4)2SO4,在培养第4d补入2.0g/LMgSO4,在培养第5d补入2.0g/LMnSO4。培养10d,菌体生物量达17.065g/L,油脂产量达6.530g/L,GLA%达23.5157%,GLA含量达1535.58mg/L,与补料前相比,生物量、油脂量、GLA%、GLA含量分别提高79.67%、217.30%、14.85%和264.43%。     

重组大肠杆菌产谷氨酰胺转氨酶培养基及发酵条件优化

对已构建好的表达谷氨酰胺转氨酶的大肠杆菌Rosetta DE3的摇瓶培养条件及发酵条件进行优化,所获优化培养基配方为葡萄糖4.0g/L,酵母膏3.0g/L,NH4Cl 4.0g/L,Na2HPO42.0g/L,K2HPO41.0g/L,MgSO4.7H2O 1.0g/L,NaCl 3.0g/L。得菌株发酵培养的最佳优化条件为装液量为25mL,接种量为5%,加IPTG浓度为0.6mmol/L,诱导时间为4h。     

利用氨基酸工业废水生产饲用产朊假丝酵母

以毛发水解法提取胱氨酸后的废液为氮源,生产饲用产朊假丝酵母。研究选取了12种饲用菌进行耐盐性驯化筛选,并对高耐盐菌株进行培养基及培养条件优化。结果产朊假丝酵母耐盐性最高,其最佳培养条件为：培养时间3 d,装液量50 ml/250 ml三角瓶,初始pH5.5。最优培养基配方为：氨基酸废水330 ml/L,玉米淀粉酶解液110 g/L,K₂>HPO₄ 20 g/L,酵母粉9 g/L,FeSO₄ 2 g/L。在此条件下,菌体得率达4.14%,较优化前提高了128.7%。     

GC/MS法同时测定电子烟烟气释放物中的5种醇类化合物

为了准确测定电子烟烟气释放物中醇类化合物的量,以1,4-丁二醇为内标建立了同时检测电子烟烟气释放物中1,2-丙二醇、1,3-丁二醇、丙三醇、二甘醇和三甘醇的气相色谱/质谱(GC/MS)方法,采用该法测定了16种电子烟样品。结果表明:①该法的检出限和定量限范围在0.08~0.32μg/口和0.27~1.07μg/口之间;回收率在90.0%~99.5%之间,相对标准偏差(RSD)为1.20%~4.93%。②所测样品中均检出1,2-丙二醇、丙三醇,均未检出二甘醇和三甘醇;4个样品中检测出1,3-丁二醇。该法灵敏度高、定性准确,适合电子烟样品烟气释放物中1,2-丙二醇、1,3-丁二醇、丙三醇、二甘醇和三甘醇的定性定量分析。      

复合诱变选育短梗霉多糖高产菌株

以本实验室保存的出芽短梗霉AP8为出发菌株,采用甲基磺酸乙酯(EMS)和紫外线(UV)复合诱变,在EMS终浓度0.4mol/L、作用时间40～60min和30W的紫外灯照射距离30cm、照射时间1.5～2.5min条件下诱变效果好,获得一株稳定遗传的多糖产量高、色素含量低的出芽短梗霉突变株UV60,产量为22.1g/L,对菌落特征和发酵特征的比较发现,突变株UV60在菌落颜色、大小、质地和发酵特性上均明显优于出发菌株AP8。通过对变异株培养基碳氮比和培养基的组成进行单因素和正交试验,其最佳摇瓶发酵培养基组成为：蔗糖50.0g/L、酵母膏1.5g/L、NaCl 1.5g/L、MgSO4 0.3g/L、K2HPO4 2.0g/L、(NH4)2SO4 0.7g/L。采用上述优化的发酵培养基,突变株UV60获得的短梗霉多糖产量为27.24g/L,多糖转化率达54.48%。     

涡旋辅助分散液液微萃取-气相色谱法测定清香型白酒中5种高级醇

采用涡旋辅助分散液液微萃取（DLLME）-气相色谱（GC）法测定清香型白酒中5种高级醇的含量。结果表明,最佳液液微萃取的提取条件为萃取剂二氯甲烷60 μL、分散剂丙酮100 μL、样品pH值5.5、酒精度15%vol、NaCl质量浓度0.19 g/mL、萃取时间30 s。在此优化条件下,正丙醇和异丁醇在含量为1.00～40.00 mg/L、异戊醇在含量为1.00～150.00 mg/L、2,3-丁二醇在含量为1.25～50.00 mg/L和β-苯乙醇在含量为0.25～10.00 mg/L的范围内具有较好的线性关系（R2＞0.99）;检出限分别为0.03 mg/L、0.01 mg/L、0.01 mg/L、0.02 mg/L和0.02 mg/L;精密度试验结果相对标准偏差（RSDs）＜7.0%,回收率为83.1%～108.3%。6种清香型白酒中正丙醇含量范围为0.064～0.116 g/L、异丁醇含量范围为0.057～0.127 g/L、异戊醇含量范围为0.262～0.450 g/L、2,3-丁二醇含量范围为0.017～0.035 g/L、β-苯乙醇含量范围为0.008～0.012 g/L。     

Metabolic effects of intraruminal administration of 1,3-butanediol or tributyrin in lactating goats

Acute effects of dietary 1,3-butanediol and tributyrin on concentrations of glucose, beta-hydroxybutyrate, and insulin in plasma were compared in lactating goats. In Experiment 1, glucose was decreased by intraruminal administration of 75 or 150 g of 1,3-butanediol or by 84 or 168 g of tributyrin. Tributyrin caused transient hyperglycemia immediately after administration. beta-Hydroxybutyrate was increased in a dose-dependent manner by tributyrin and increased independently of dose by 1,3-butanediol. Tributyrin, but not 1,3-butanediol. Tributyrin, but not 1,3-butanediol, caused large increases of insulin in plasma. In Experiment 2, 75 g of 1,3-butanediol or 84 g of tributyrin administered intraruminally decreased glucose, whereas 73 g of butyric acid (pH 5.6) increased glucose compared with water or 25.6 g of glycerol. All treatments produced transient hyperglycemia immediately after administration. Tributyrin, butyric acid, or 1,3-butanediol greatly increased beta-hydroxybutyrate compared with administration of water or glycerol. Concentrations of beta-hydroxybutyrate in both experiments increased more slowly for 1,3-butanediol than for tributyrin. Both 1,3-butanediol and tributyrin decreased glucose and increased beta-hydroxybutyrate. Because 1,3-butanediol does not stimulate increases of insulin in plasma, it may be more desirable than tributyrin for inducing metabolic changes characteristic of lactation ketosis.     

茅台酱香型酒糟中总黄酮及总多酚含量的测定

建立酱香型酒糟中总黄酮及总多酚含量测定方法。以槲皮素为对照品,采用紫外分光光度法测定酱香型酒糟中总黄酮含量;以没食子酸为对照品,用Folin-Ciocateu比色法测定酒糟中总多酚含量,并对其测定条件进行了优化。结果表明,Folin-Ciocalteu比色法最优的测定条件为：加入Folin-Ciocalteu试剂2.5 mL、10% Na2CO3溶液4 mL,在室温下显色80 min。槲皮素和没食子酸质量浓度分别在0.021 0～0.105 0 g/L(R2 = 0.997 6)和0.000 5～0.005 7 g/L(R2 = 0.999 7)范围内与吸光度值线性关系良好,平均回收率分别为99.12%、99.82%,相对标准偏差(RSD)分别为0.61%、0.33%;茅台酱香型酒糟中总黄酮及总多酚含量分别为20.40 mg/g和4.78 mg/g。此方法操作简单,稳定性以及重复性良好,结果准确可靠,适用于酱香型酒糟总黄酮和总多酚含量的测定。     

类球红细菌产辅酶Q_(10)发酵工艺的优化

类球红细菌(Rhodobacter sphaeroides)是一类高产辅酶Q10的光合细菌,该研究对R.sphaeroides EIM-8产辅酶Q10的发酵条件和培养基进行了优化。采用单因素结合响应面设计优化后的培养基组成为:葡萄糖27.8 g/L、(NH4)2SO4 4.9 g/L、谷氨酸4.7 g/L、玉米浆粉2.5 g/L、MgSO49.5 g/L、FeSO41.5 g/L,NaCl 3.5 g/L,KH2PO44.0 g/L、辅液15.0 mL/L;通过单因素试验方法确定的培养条件为:pH 6.5、温度32℃、接种量10%、装液量40mL/250 mL,此条件下辅酶Q10产量可达128.9 mg/L,比优化前提高了89.0%。     

热带假丝酵母木糖还原酶在酿酒酵母细胞表面展示

利用酿酒酵母(Saccharomyces cerevisiae)表面展示系统,将来源于热带假丝酵母(Candida tropicalis)的木糖还原酶基因xyl1嵌入带有His-Tag的酿酒酵母α-凝集素展示载体pICAS-His,构建重组质粒pICAS- His-Ctxyl1,并转化到酿酒酵母宿主菌酿酒酵母MT8—1,通过流式细胞仪快速检测和筛选,得到重组菌株MT8- 1/pICAS-His—Ctxyl1。将重组酵母用于葡萄糖(15g/L)和木糖(5g/L)的混合糖发酵研究,结果表明,重组酿酒酵母MT8/1/pICAS-His—Ctxyl1细胞具有良好的生长和产酶特性,同时能转化木糖生产木糖醇,在培养基中2.5g/ L木糖转化生成2.5g/L木糖醇,转化率达98.7%。     

Metabolic responses of lactating goats to feed restriction and dietary 1,3-butanediol

Feed restriction and dietary 1,3-butanediol were used with lactating goats in an attempt to induce metabolic changes characteristic of bovine lactation ketosis and fatty liver. In Experiment 1, midlactation goats were fed 80, 102, or 114% of metabolizable energy requirements and 0, 50, or 100 g/d of 1,3-butanediol. Concentration of beta-hydroxybutyrate in blood plasma decreased with increasing metabolizable energy but was increased greatly at 2 h after goats were fed 50 or 100 g butanediol and remained elevated at 6 h postfeeding with 100 g of butanediol. Concentration of glucose in plasma was decreased at 2 and 6 h postfeeding in goats fed 100 g of butanediol. In Experiment 2, goats in early lactation were fed for ad libitum intake or were restricted to 70% of ad libitum intake with 1,3-butanediol included at 10% of diet DM. The treatment decreased milk production, increased concentrations of beta-hydroxybutyrate and nonesterified fatty acids, and decreased the concentration of insulin and the insulin to glucagon ratio in plasma. Concentrations of glucose, acetate, and glucagon in plasma were not affected. After 28 d of treatment, concentration of total lipid in liver was increased, but concentrations of glycogen and triglyceride were unaffected. Changes caused in goats by feed restriction plus dietary 1,3-butanediol were characteristic of subclinical lactation ketosis in cows, but the response was more moderate than seen previously in cows.     

高产辅酶Q10类球红细菌的化学诱变筛选及其发酵培养基优化

利用菌株对（维生素K3+叠氮化钠+对羟基苯甲酸）的复合抗性作为筛选标记,对产辅酶Q10的类球红细菌（Rhodobacter sphaeroides）使用亚硝基胍进行化学诱变,筛选高产菌株并采用响应面法优化其发酵培养基。结果表明,经诱变选育得到一株遗传稳定的辅酶Q10高产菌株R.sp3-7,其最佳发酵培养基组成为：葡萄糖31.7 g/L,玉米浆干粉5.6 g/L,（NH4）2SO4 5.3 g/L、谷氨酸钠 3.0 g/L、NaCl 3.0 g/L、MgSO4·7H2O 12.5 g/L、KH2PO4 3.0 g/L、CaCO3 2.0 g/L、辅液1 mL/L。在此培养条件下,诱变菌株R.sp3-7的辅酶Q10产量达（93.63±0.59） mg/L,与出发菌株相比提高了116.8%。 关键词：中图分类号：TS201.3 文章编号：0254-5071（20１6）06-0090-06 doi:     

Physiological effects of 1,3-butanediol fed to cattle.

In four trials with growing cattle we observed effects of 1,3-butanediol on rumen fermentation end products, blood components, growth rates, feed efficiency, and body composition. Diets A and C contained 80% grain: 20% alfalfa pellets and 40% grain: 60% alfalfa pellets; in diets B and D, 1,3-butanediol replaced 4% of diets A and C. Feeding 4% 1,3-butanediol caused no significant differences in rumen pH, volatile fatty acid ratios, blood glucose, or blood ketones. Cattle fed 4% 1,3-butanediol had rates of gain and feed efficiency equivalent to and often better than cattle fed the same diet without butanediol. Body composition was not affected by feeding 4% 1,3-butanediol. Results indicate 4% 1,3-butanediol can be fed to cattle without adverse physiological effects and may improve performance. When 6% or more 1,3-butanediol was fed, blood ketones were elevated considerably, animals were hyperactive, gained less weight, and feed efficiency was lower.