一株高产葡聚糖肠膜状明串珠菌肠膜亚种的筛选及发酵条件优化

为拓宽高产葡聚糖的种质资源,以四川泡菜为分离源从中筛选高产葡聚糖的乳酸菌菌株,采用苯酚硫酸法对筛选菌株进行葡聚糖产量测定,并对菌株进行生理生化及16S rRNA基因和rpo B看家基因序列的同源性分析鉴定和高产葡聚糖发酵条件优化。结果表明:菌株LM187产葡聚糖能力较强,达到38.24 g/L,进一步鉴定为肠膜状明串珠菌肠膜亚种(Leuconostoc mesenteroides subsp. mesenteroides);采用响应面中心组合实验优化产葡聚糖发酵条件,得到最优培养条件为发酵时间35 h、发酵温度30℃、蔗糖质量分数21%和接种量3%。在此优化条件下,葡聚糖产量为56.54 g/L,较优化前提高了1.48倍。肠膜状明串珠菌肠膜亚种LM187具有高产葡聚糖的能力,可作为开发保健型泡菜的潜在优质功能性微生物资源。     

耐热β-葡聚糖酶产生菌AS35产酶条件的研究

本试验对枯草芽孢杆菌(Bacillus subtilis)AS35产β-葡聚糖酶的发酵条件进行优化研究。单因素实验结果表明:该菌产β-葡聚糖酶的最佳碳源为麦糟粉,添加量为3.0%(w/v),最佳麦糟粉颗粒直径为0.5mm左右;最佳氮源为蛋白胨,添加量为0.8%(w/v);最佳培养基初始pH为7.0;最佳摇瓶装液量为50mL/250mL三角瓶;最佳发酵温度为36℃。在上述优化条件下,以接种量10%(v/v)接种菌龄18h的种子于发酵培养基中,200r/min摇床培养60h,β-葡聚糖酶酶活达到32.5U/mL。同时,耐热性实验表明,该酶最适反应温度为65℃。     

β-葡聚糖酶发酵培养基的优化研究

为了对芽孢杆菌发酵产β-葡聚糖酶的培养基进行进一步的研究,以芽孢杆菌为发酵菌,通过单因素试验和三因素三水平正交试验法对芽孢杆菌的10L发酵罐发酵产酶培养基进行优化。实验结果表明:芽孢杆菌产β-葡聚糖酶的最优培养基为甘油含量6g/L、酵母粉含量24g/L和NaCl含量10g/L;发酵条件:接种量为10%、初始发酵pH值为7、培养温度为37℃、转速为350~950r/min、通风量为1vvm和发酵时间15h。经过3批发酵实验验证,最优条件下β-葡聚糖酶最高酶活可达3112U/mL。     

液态深层发酵型木瓜果醋的发酵生产研究

本研究以云南药木瓜为原料,沪酿1.41醋酸菌为菌种,液态深层发酵酿制木瓜果醋.研究分析了温度、醋酸菌接种量、初始酒度、初始酸度、通风量对产酸发酵的影响,确定了木瓜果醋酿制的最佳工艺参数为温度35℃、接种量11%(V/V)、初始酒度4.7%(V/V)、初始酸度1.8g/100ml(以醋酸计)、通风量200r/min(以摇床转速计),最终酸度达到4.96g/100ml,发酵率为67.23%.在最佳工艺条件下发酵,醋酸菌在发酵第6d达到对数生长期.酿制出来的木瓜果醋颜色金黄、澄清透亮,醋味浓郁,同时具有木瓜的特殊清香味.     

产骨胶原蛋白酶菌的筛选及发酵条件优化的研究

经初筛和复筛从骨骼厂土样中筛选出高产骨胶原蛋白酶的菌种MBL1,针对其产酶的最佳菌龄和发酵条件进行优化研究.结果表明,最佳接种菌的种龄选用培养24h,接种量为5%,起始pH值为7.0,150ml三角瓶最佳装液量20ml,发酵温度32℃.且在优化后的发酵条件下,该菌产酶酶活较优化前酶活提高了35%,达到49.83U/ml.     

米酒乳杆菌素分离纯化条件研究

目的：对米酒乳杆菌产生的广谱细菌素进行分离纯化技术研究。方法和结果：采用活性炭脱色、冷乙醇沉淀、Sephadex G50 葡聚糖凝胶层析和高效液相色谱技术对米酒乳杆菌产生的细菌素进行分离纯化。结果表明,发酵浓缩液脱色的最佳工艺条件为活性炭加量3.5%、温度40℃、脱色时间20h,脱色率达64.5%。冷乙醇沉淀的最佳乙醇终浓度为80%,葡聚糖凝胶层析的洗脱液流速为0.5ml/min,上样量3ml,高效液相色谱纯化的流动相为90% 的甲醇水溶液,在此条件下可得到电泳纯的细菌素。结论：确定了米酒乳杆菌素的分离纯化条件,为细菌素的理化性质和抑菌机理的研究提供依据。     

产葡聚糖明串珠菌的分离鉴定及发酵条件研究

从实验室保存的开菲尔粒中分离筛选高产葡聚糖菌株.经乳酸菌的初筛和胞外多糖产糖量分析,筛选出一株能高产葡聚糖的菌株,命名T-L1,结合形态学特征和16S rRNA的序列分析,将该菌株鉴定为明串珠菌(Leuconostoc mesenteroides),采用单因素和正交试验对该菌发酵条件进行优化研究,结果表明该菌的最佳发酵条件发酵温度为28℃、发酵时间为24 h、接种量体积分数为5％,培养基起始pH值为6.5.在此优化条件下,葡聚糖的产量为30.12 g/L,比优化前提高了41％.     

两种益生菌发酵促使牛骨粉中钙转化的比较研究

选用鼠李糖乳杆菌、婴儿双歧杆菌发酵牛骨粉。采用L9(34)正交试验设计,对蔗糖添加量、接种量、骨粉浓度、发酵时间进行筛选。以游离钙转化率为特征性指标探讨发酵条件对游离钙转化率影响的变化规律,并且比较两种益生菌在体外模拟胃肠环境中的活菌数。结果表明：鼠李糖乳杆菌的最适发酵条件为蔗糖添加量11%、接种量3%、骨粉浓度5g/100ml、发酵时间72h,婴儿双歧杆菌的最适发酵条件为蔗糖添加量8%,接种量5%、骨粉浓度10g/100ml、发酵时间72h;鼠李糖乳杆菌对牛骨粉中钙的转化率达32.0%,约为婴儿双歧杆菌的3倍;发酵时间是影响游离钙转化率的重要因素;鼠李糖乳杆菌活在模拟胃、肠液中的菌率是分别婴儿双歧杆菌的10.38、16.35倍。     

粗壮脉纹胞菌复合多菌种发酵茶粕产纤维素酶的研究

用粗壮脉纹胞菌分别复合东方伊莎酵母、里氏木霉、绿色木霉、乳酸杆菌固态发酵已去除茶皂素的茶粕,通过测定发酵产物中3种纤维素酶：外切葡聚糖酶（C1）、内切葡聚糖酶（Cx）、β-葡萄糖苷酶（β-G）及总酶滤纸酶（filter paper activity,FPA）的酶活力来探讨其分解粗纤维素的协同作用。粗壮脉纹胞菌和绿色木霉混合发酵产生的C1酶酶活力较粗壮脉纹胞菌单菌发酵提高了51.09%,粗壮脉纹胞菌和绿色木霉复合发酵较单菌发酵延长了其纤维素酶分泌的周期,96 h时FPA酶活力达到2.782 U/g;粗壮脉纹胞菌复合里氏木霉、绿色木霉混合发酵组在发酵10 d后对茶粕粗纤维的最终降解率分别达到了64.19%和61.59%;接种量对单菌和混合菌发酵产纤维素酶影响总体趋势是随着接种量增加酶活力提高,但粗壮脉纹胞菌单菌发酵纤维素酶酶活力在接种量超过9 mL/100 g后开始下降。表明粗壮脉纹胞菌复合里氏木霉、绿色木霉混合发酵降解纤维素具有协同作用。     

玉米淀粉和黄浆发酵罗耳阿太菌双响应值优化

为获得罗耳阿太菌β-1,3葡聚糖酶和胞外多糖同时高产的发酵条件,以玉米淀粉和玉米黄浆作为发酵培养基的重要组分,以罗耳阿太菌β-1,3葡聚糖酶产量和胞外多糖产量为指标,选择培养温度、培养时间、摇床转速为优化因素,在单因素试验的基础上,通过响应面试验进行双响应值优化,对所得结果的三维图和等高线叠加图进行分析,获得了双指标同时达到最优的发酵条件。结果表明:接种量5%、培养温度28.5℃、培养时间7.5 d、摇床转速180 r/min时,粗酶产量39.96 U/m L,多糖产量18.11 g/L,分别达到了预测值的98.96%和99.27%。     

Physicochemical Properties and Phytochemical Components of Spiced Cucumber-Pineapple Fruit Drink

A blend of equal portions of cucumber (50%) and pineapple (50%) juices was mixed with clove and ginger powder spices at 0.25% (CPCLG1), 0.5 (CPCLG2), 0.75% (CPCLG3) and 1% (CPCLG4) (w/v) respectively to develop a new fruit drink with health benefits. The juice blend without spice extract (CP) was used as reference sample. Samples were analysed for oBrix, pH, acidity, specific gravity, total solids and ascorbic acid (vitamin C). The phytochemicals present in the products were also determined using chemical method and Gas chromatography-mass spectrometry (GC-MS). There was significant reduction in the oBrix from 8.08 to 7.60%, pH (4.41 to 4.36) and total solid (7.96 to 7.71%). Specific gravity was 1.020 while the acidity increased from 0.20 to 0.22% as the levels of spices increased. The phytochemical compounds identified in the products include alkaloids, flavonoids, saponins, steroids, tannins, terpenoids and phlobatannins. The quantitative phytochemical contents showed 0.1 μg/ml Isoquinoline in CP, 0.03271 μg/ml Isoquinoline in CPCLG1; 0.00199 μg/ml Octanoid acid, 0.00224 μg/ml Metroprolol, 0.00231 μg/ml Fumaric acid, 0.00263 μg/ml Benzoquinone, 0.00264 μg/ml Betaxolol, and 0.002471 μg/ml 1-nonene in CPCLG2; 0.00070 μg/ml Limonene and 0.00146 μg/ml Caryophyllene in CPCLG3 and 0.00048 μg/ml P-Benzoquinone, 0.00074 μg/ml 4H-Quinolizine, 0.00017 μg/ml 3-pyridinepropanol, 0.00559 μg/ml Chlorogenic acid, 0.00559 μg/ml Camphene and 0.00089 μg/ml Benzofuranone in CPCLG4. The types and quantities of phytochemicals present in the products were influenced by the formulation of the products.     

高产SOD酵母菌的诱变选育及发酵条件研究

用紫外线诱变酿酒酵母B094原生质体，得到1株SOD含量较高的菌株BUV094，其SOD含量为720U／g湿菌体。用正交试验及单因了条件试验对该突变株进行培养基组成及发酵条件的研究，结果表明：其最佳培养基组成为葡萄糖2％，蛋白胨1％，酵母膏1％，7Be麦芽汁9％（V／V），CuSO475mmol／ml，ZnSO425mmol／ml；最佳培养条件为：初始pH5．5，摇瓶装量为75ml／500ml三角瓶，接种量为15％，培养时间24h。在此条件下，该菌细胞生物量为5．39g／100ml，SOD含量为1024U/g湿菌体，SOD产量为5520U／100ml，SOD产量较出发菌株提高103％。     

薏苡仁在曲霉发酵过程中成分及抗氧化性变化的研究

研究以黑曲、米曲、红曲分别发酵薏苡仁,然后对发酵液中多肽、总糖、多酚、DPPH 自由基清除率进行测定,以考察不同曲霉发酵过程中,薏苡仁发酵液中的各种物质含量及抗氧化性变化。结果表明：发酵过程中多肽含量、总酚含量、蛋白质水解度、DPPH 自由基清除率都随着发酵时间的延长而不断增加。以黑曲霉发酵时各指标分别达到6.14mg/ml、0.10mg/ml、24.9%、10.2%;以米曲霉发酵时各指标分别达到2.58mg/ml、0.089mg/ml、31.1%、10.4%;以红曲霉发酵时各指标分别为2.93mg/ml、0.086mg/ml、14.7%、18.9%。发酵过程中总糖的含量黑曲霉组、米曲霉组均上升,各达到84.0mg/ml、76.1mg/ml;而红曲霉发酵后总糖含量下降明显,降至8.5mg/ml。     

山羊初乳成分及其免疫球蛋白构成变化的研究

动物初乳对其后代健康的作用已经得到了广泛的证实,其对人类健康的影响也受到了越来越多的重视.本研究探讨了阿尔卑斯山羊产后一周内乳成分和其中免疫球蛋白(Ig)含量的变化.结果表明:产后48h内山羊奶的蛋白质、总固形物和免疫球蛋白含量均快速下降;脂肪含量在前三次的初乳中逐渐升高,随后又缓慢下降;乳糖含量保持相对稳定于4%～5%;经产山羊前三次初乳的蛋白含量分别达到了16.5%、10.7%和6.8%,显著地高于初产山羊前三次初乳的蛋白含量(7.0%、4.8%和4.1%);经产山羊前两次初乳的IgG含量达到了59.9mg/ml和34.7mg/ml,IgM含量达到了6.14mg/ml和2.37mg/ml,显著地高于初产山羊前两次初乳的IgG含量(18.1mg/ml和13.9mg/ml)和IgM含量(1.95mg/ml和0.98mg/m1);经产和初产山羊初乳的IgA含量无显著差异.     

枸杞果醋饮料的配方优化及品质分析

以枸杞为原料制备枸杞汁、枸杞醋,并添加白砂糖及菊花提取液制备枸杞果醋饮料,通过单因素试验及正交试验优化枸杞果醋饮料配方,并对其感官、理化及微生物指标进行分析。结果表明,枸杞果醋饮料的最佳配方为：枸杞醋8%,枸杞汁15%,白砂糖6%,菊花提取液15%。在最优配方条件下,枸杞果醋饮料感官评分为95.47分,枸杞果醋饮料口感柔和、酸甜爽口。总酸、不挥发酸及氨基酸态氮含量分别为0.66 g/100 m L、0.36 g/100 m L、0.07 g/100 m L,总糖、蛋白质、脂肪含量分别为9.62 g/100 m L、0.48 g/100 m L、0.06 g/100 m L,其理化及微生物指标均符合国家农业部标准NY/T 2987—2016《绿色食品果醋饮料》要求。     

白酒生产用己酸菌发酵液发酵条件及培养基组成的优化

以白酒生产用的己酸菌为研究对象,对其发酵条件和培养基的组成进行了优化。 首先采用单因素试验优化了己酸菌的发酵 条件,随后采用Plackett-Burman试验、最陡爬坡试验、响应面法优化了己酸菌发酵培养基的组成。 结果表明,最佳发酵条件为发酵温度 33 ℃、接种量5%、装液量50 mL/100 mL、发酵时间11 d。 最佳发酵培养基的组成为乙酸钠1.644 g/100 mL、硫酸铵1.213 g/100 mL、生物 素0.013 g/100 mL、碳酸钙1.0 g/100 mL、磷酸氢二钾0.025 g/100 mL、硫酸镁0.025 g/100 mL、酵母浸粉0.625 g/100 mL、乙醇2.5 mL/100 mL、 对氨基苯甲酸0.062 5 g/100 mL。 在此优化培养工艺下,获得的己酸菌发酵液中己酸产量达18.93 g/L,比初始发酵工艺的发酵液（7.03 g/L） 提高了169%。     

Motility and fertility of frozen bull spermatozoa in tris-yolk and milk extenders containing amikacin sulfate

Amikacin sulfate, an aminoglycoside antibiotic, effective against some streptomycin-resistant organisms, was tested in bull semen extended with whole milk and egg yolk-Tris extenders in a 6 X 5 X 2 factorial arrangement. Semen from six bulls was frozen in extenders containing 500, 1000, and 2500 microgram/ml of amikacin, a control containing 500 IU/ml of penicillin plus 1000 micrograms/ml of streptomycin plus 500 IU/ml of polymyxin, and control plus amikacin (1000 micrograms/ml). Semen was frozen in .5 ml polyvinyl chloride straws in liquid nitrogen vapor and stored in liquid nitrogen for 2 wk. Straws were thawed in a water bath at 35 degrees C for 30 s. Postthaw motility of spermatozoa in each treatment was assessed using two straws by two observers independently. At amikacin concentrations of 500, 1000, or 2500 micrograms/ml, the percentage of motile spermatozoa averaged 47, 45, and 36% in whole milk and 52, 46, and 46% in egg yolk-Tris. Control and control plus amikacin (1000 micrograms/ml) were 49 and 47% in whole milk and 48 and 52% in egg yolk-Tris. Only amikacin in milk at 2500 micrograms/ml, depressed sperm motility. A fertility trial was conducted to compare the control mixture of antibiotics in whole milk with whole milk also containing 500 micrograms/ml of amikacin. The nonreturn rate to 1182 first service inseminations with semen treated with amikacin (500 micrograms/ml) plus control antibiotics was 70.4% compared with 70.5% for 1494 first service inseminations with control semen. Amikacin can safely be added to whole milk and egg yolk-tris extenders to control streptomycin-resistant organisms.     

Herd and state means for somatic cell count from dairy herd improvement

To determine the impact of reducing the current legal limit of 750,000 cells/ml for somatic cell count (SCC) in US market milk, data were examined from 539,577 herd test days for Dairy Herd Improvement herds on test during 1996 and 1997. Somatic cell scores for individual cows were converted to SCC. The SCC for each cow was weighted by milk yield and used to compute herd mean on test day. The mean for each state was derived by weighting herd test-day SCC by herd test-day milk yield. State means were lowest in the West and highest in the Southeast. The percentage of herd test days with an SCC of >750,000 cells/ml ranged from 0 to 14% across states; the mean was 4%. Only 1% of the herd tests were >750,000 cells/ml on 2 consecutive test days. Mean SCC in the United States was 307,100 cells/ml for 1996 and 313,500 cells/ml for 1997. Mean SCC was lower during October through January (280,000 to 300,000 cells/ml) than during July and August (340,000 cells/ml). Herd size and SCC were negatively related; larger herds had lower SCC. Because records of some cows treated with antibiotics were included in the data, herd SCC means likely were higher than corresponding bulk tank SCC. Most herds had test-day SCC that were substantially below legal bulk tank limits and could have met lower limits (e.g., 500,000 cells/ml).     

一株纤维素酶真菌的筛选鉴定及产酶条件优化

以羧甲基纤维素钠为唯一碳源,从自然界筛选出一株纤维素酶高产菌HJC-79,通过单因素试验和正交试验对其培养基成分和发酵条件进行优化。结果表明,经形态观察和ITS序列分析,菌株HJC-79被鉴定为杂色曲霉（Aspergillus versicolor）。最佳培养基成分为蔗渣、麸皮和Mandels营养盐液的添加量分别为2%、9%、89%;最佳发酵条件为发酵温度33 ℃,初始pH值为4,接种量9.0%,发酵时间48 h。在此优化条件下,葡聚糖内切酶、β-葡萄糖苷酶和滤纸酶酶活分别为2.22 U/mL、1.93 U/mL、1.53 U/mL,分别比优化前提高了125.55%、76.37%和172.47%。     

桤叶唐棣果醋发酵工艺优化及有机酸和挥发性成分分析

以桤叶唐棣为原料,经半固态发酵酿制桤叶唐棣果醋,以总酸作为评价指标,优化桤叶唐棣果醋发酵工艺,并对发酵前后有机酸和挥发性成分进行比较。结果表明,最佳发酵条件为初始酒精度9%vol,果酒与果渣质量比1.0:1.6,醋酸菌添加量2.0%,在30 ℃条件下发酵18 d,测得桤叶唐棣果醋总酸含量为4.75 g/100 mL。桤叶唐棣果醋中以醋酸（5.00 mg/mL）、丙二酸（0.41 mg/mL）、乳酸（0.25 mg/mL）为主。主要挥发性物质为3-羟基-2-丁酮（15.86%）、乙酸（35.62%）、乙酸乙酯（24.32%）、苯甲醇（17.60%）、苯甲酸（6.60%）。