冰核活性细菌发酵培养基的优化筛选

研究了不同营养构成及添加不同营养物质对冰核细菌生长和冰核活性的影响 ,采用正交试验优化了培养基组成。优化后的培养基最佳配方为 :在MPDA培养基中添加 0 4%半乳糖、1 5 %酵母粉、0 0 1%粗磷脂。在优化后的培养基中培养供试菌 ,菌体收获量 (干重 )比在MPDA培养基中培养增加 2 4g/L ,冰核活性增加 2 4 3 %。     

Xanthomonas ampelina发酵培养基组成的优化

本文采用正交实验的方法优化了Xanthomonas ampelina TS206发酵培养基的配方,从而达到了提高冰核活性细菌发酵所得的生物量和细菌的冰核活性的目的。实验结果表明较优化的培养基配方为:牛肉膏3.0g/L、蛋白胨5.0g/L、甘油25.0g/L、蔗糖20.0g/L、山梨醇2.0g/L、酵母粉20.0g/L、玉米浆20.0g/L、K2HPO4 1.5g/L、pH=7.0;最终发酵液的生物量达到8.2g/L(干重),冻滴率达到81%。     

冰核活性细菌Xanthomonas ampelinaTS206实验室水平发酵生产工艺条件优化的研究

优化冰核活性细菌XanthomonasampelinaTS206实验室水平发酵生产的工艺条件为将冰核活性细菌应用在工业化生产提供了重要的技术资料。本文对冰核活性细菌XanthomonasampelinaTS206实验室水平的发酵生产工艺条件进行了系统的研究,结果表明这种冰核活性细菌的最适培养条件为:培养温度18℃,装液量40ml/250ml,摇床转速180r/min,接种量3%,发酵液的初始pH8。实验还确定了冰核活性细菌在发酵48h时达到对数生长期。采用上述发酵条件培养XanthomonasampelinaTS206可以使细菌在冰核活性不降低的条件下发酵液的菌体浓度达到2.87×1010个/ml,菌体干重增加了37.7%。     

植物抽取物对冰核活性细菌表达冰核活性蛋白的影响

植物提取物对于冰核活性细菌表达冰核活性蛋白具有明显的促进作用。通过添加适量的芥菜籽热水抽取物培养冰核活性细菌 (XanthomonasampelinaTS2 0 6) ,细菌的冰核活性比原来提高了 37.5 %,菌体的生物量提高了 36.2 %。芥菜籽抽提物作为一种效应物分子激活启动子 ,在冰核蛋白基因的表达中起到了重要作用 ,从而显著地提高了细菌高水平表达冰核活性蛋白的能力。     

冰核活性细菌发酵生产冰核活性蛋白产物的研究进展

对国内外关于冰核活性细菌 (简称INA细菌 )发酵生产冰核活性蛋白的条件 ,发酵过程的调控手段 ,及其对发酵生产的相关技术方法进行了较为详尽的综述 ,为研究开发这一新型蛋白资源提供较为全面的背景材料     

葡萄叶片上冰核细菌的分离鉴定

为了检测葡萄上冰核活性细菌的分布情况,并探究其对葡萄叶片霜冻害的影响,本试验对葡萄叶片上的冰核活性细菌进行了分离、鉴定和冰核活性的测定,并检验了活性较高的菌株对叶片冻害的效果。试验通过分离共获得10株冰核活性细菌,活性较高的1、2号菌株都是假单孢菌属(Pseudomonas)。用1、2号菌处理葡萄叶片,进行霜冻胁迫,叶片的霜冻指数分别达到83.67%和90.42%,而对照组为66.13%;活性最好的2号菌可以提高叶片过冷点2.3℃。     

培养基组成对冰核生产菌Xanthomonas ampelina TS206产胶率的影响

培养基中不同的碳源、氮源和无机盐影响冰核细菌的生长、冰核蛋白的表达及黄原胶的产率。蔗糖、麦芽糖和山梨醇均有利于冰核蛋白的生产,而且麦芽糖和山梨醇的产胶率要明显低于其它碳源;蛋白胨和玉米浆均可以作为冰核蛋白生产的优良氮源使用,无机氮不适合作为唯一氮源使用,但适量的KNO3和蛋白胨结合使用时,可以促进菌体的生长和冰核活性的表达,但同时也促进了黄原胶的形成;适量地添加无机盐对菌体的生长和冰核活性的表达有一定的促进作用,对黄原胶的合成没有明显的抑制作用。     

降低冰核生产菌X．ampelina TS206黄原胶产率的研究

培养基中不同的碳源影响冰核细菌的冰核活性、生物量和产胶率，麦芽糖、山梨醇、蔗糖等均可做为促进冰核细菌生长碳源，麦芽糖作为碳源加入发酵培养基中生物活性和生物量均较高，同时产胶率低于葡萄糖和蔗糖十倍左右。麦芽汁代替麦芽糖可以降低产胶率，但效果不如麦芽糖。在培养基中加入0．1ml，100ml乳酸可以有效地抑制黄原胶的产生，而对冰核活性和生物量影响不大。     

冰核活性细菌灭活方法的初步研究

探讨了几种灭活方法对冰核活性细菌死亡率和冰核活性的影响。结果表明:超声波破碎法使冰核细菌10025A的死亡率达到98%以上,但冰核活性有所下降。辐射处理可使冰核细菌达到100%的死亡率,不同菌种对辐射的敏感程度不同,冰核活性的变化也有所不同。其它方法处理的冰核细菌死亡率较低。     

冰核活性细菌Erwinia herbicola发酵培养条件的研究

研究了不同培养条件对冰核活性细菌冰核活性和菌体生物量的影响。结果表明：培养时间、温度、低温诱导和培养基pH值对培养液中冰核活性细菌的总活性有不同的影响。研究确定的最适发酵条件组合为培养温度18℃，培养基pH6．5，培养时间32h。     

海藻酸钙固定化对Xanthomonas campestris206冰核活性的影响

应用正交法研究了海藻酸钙固定化对Xanthomonascampestris 2 0 6冰核活性的影响。结果表明 ,对固定化小球冰核活性影响程度大小的顺序依次为 :菌悬液用量 >海藻酸钠用量 >CaCl2 浓度 >固化时间 ,而对渗漏量影响程度大小的顺序依次为 :菌悬液用量 >固化时间 >CaCl2 浓度 >海藻酸钠用量。研究中还发现 ,将冰核活性细菌固定化在热稳定性方面实际意义不大。     

突变型黄单胞杆菌产黄原胶条件的优化

该文对从甘蓝腐烂根部的土壤中分离选育的黄单胞杆菌C2发酵产黄原胶的条件进行了优化。通过单因素试验对培养基（碳源、氮源及碳氮源的浓度）和发酵条件（接种量、发酵时间、温度、pH、装液量）进行优化;为提高黄原胶的胶体性能,在培养中添加不同量的游离丙酮酸。结果表明,液态发酵生产黄原胶的最佳碳源、氮源分别为蔗糖55 g/L,鱼粉10 g/L。黄单胞杆菌C2菌株在接种量为4%,pH值为7,于28 ℃、130 r/min条件下发酵60 h时,黄原胶产量达到34 g/kg。添加游离丙酮酸2 g/L时发酵液黏度为16.0 Pa·s,此时胶体性能最好。     

富含谷氨酸酿酒酵母的筛选及发酵培养基优化

从农家自酿葡萄酒中筛选出一株富含谷氨酸酿酒酵母菌（Saccharomyces cerevisiae）F-5,其26S rDNA核苷酸序列与S. cerevisiae TY12的26S rDNA核苷酸序列同源性为100%。以胞内谷氨酸含量为目标,采用响应面法对其发酵培养基进行了优化,建立糖蜜、工业蛋白胨和KH2PO4的二次回归模型,确定培养基最佳配方为：糖蜜（含30%蔗糖）100 mL/L、酵母浸粉10 g/L、工业蛋白胨20 g/L、MgSO4·7H2O 1 g/L、KH2PO4 0.5 g/L、FeSO4·7H2O 2 g/L。在此优化培养基中发酵培养24 h,胞内游离谷氨酸达到了3.29%,比优化前提高了87.8%。     

Freezing and Ice Recrystallization Properties of Sucrose Solutions Containing Ice Structuring Proteins from Cold-Acclimated Winter Wheat Grass Extract

ABSTRACT: The freezing properties of sucrose solutions containing ice structuring proteins (ISPs) from cold-acclimated winter wheat grass extract (AWWE) were evaluated. Neither significant ice nucleation nor thermal hysteresis activity in unpurified AWWE were detected ( P > 0.05). Ice recrystallization in sucrose solutions was assessed by bright field microscopy. Ice crystal growth was significantly reduced with the addition of more than 0.05% total protein from AWWE in 23% sucrose solutions frozen under static conditions and temperature cycled under long periods of time (60 min). Ice recrystallization inhibition was not evident when the samples were temperature cycled for shorter periods of time (10 min), indicating that an adsorption time may be required for the ISPs to be significantly active. The ice recrystallization was reduced with the increased on ISP concentration until reaching a plateau after adding 0.13% total protein from AWWE, representing a surface coverage of 9 mg protein/m² ice (assuming 100% adsorption). The reduction of ice crystal growth was as high as 74% compared with the control. This ice recrystallization inhibition offers significant opportunity for the use of ISPs from AWWE in frozen foods.     

Effect of stabilizers on the shelf-life of Penicillium frequentans conidia and their efficacy as a biological agent against peach brown rot

Stabilizers were added to conidia of Penicillium frequentans at two different points of the production-formulation process to improve shelf-life of conidia stored at different temperatures. Effects were also tested on conidial germination and production. Germination of conidia without additives was 90.2%; sodium chloride, potassium chloride, triton TX100, dimethyl sulphoxide, peroxidase, 0.375% and 0.075% ascorbic acid, 7.5% and 3.75% sucrose, and 7.5% and 3.75% d-sorbitol reduced significantly (P=0.05) conidial germination, while no effect was observed with glucose, lactose, maltose, sodium glutamate, glycerol, peptone, sodium alginate, carboximethylcellulose, Tween 80, and gelatine. Production of P. frequentans conidia in solid-state fermentation without additives was 1.07 conidia x 10(8) g(-1) of dry substrate. The highest tested doses of glucose, lactose, maltose, sodium glutamate, and glycerol enhanced production of P. frequentans, while the lowest tested doses of d-sorbitol and ascorbic acid reduced it. No significant effect was observed with sucrose, peptone, sodium alginate, carboximethylcellulose, gelatine and Tween 80. Conidial germinability after one year of storage at different temperatures was studied in some formulations. It was lower than 18% after 365 days of storage at room temperature in control samples (without any additive), being enhanced when 7.5% glucose, 7.5% glycerol, or 1.5% sodium alginate was added to the substrate in bags before fermentation; or when 7.5% glucose, 7.5% sodium glutamate, or 1.5% sodium alginate was added to conidia before drying. Germinability of conidia produced without any additive and stored at 4 degrees C was significantly higher (38%) than at room temperature, being enhanced when 7.5% glycerol or 1.5% sodium alginate was added to the substrate in bags before fermentation; and when 7.5% glucose, or 1.5% sodium alginate was added to conidia before drying. No effect was observed with the presence or absence of light or high vacuum. Four formulations of P. frequentans conidia reduced disease incidence by more than 55%. The relationship of the disease control with the viability of P. frequentans was discussed.     

嗜热链球菌的筛选及其高活性菌粉制备

为获得优良酸奶发酵剂嗜热链球菌菌株,并确定制备高活性菌粉的培养基与保护剂,利用改良TJA培养基,自市售酸奶中分离纯化出6株球菌,经生理生化试验鉴定为嗜热链球菌。进一步对它们的发酵特性进行比较,结果表明ST4发酵性能优良。以10%复原脱脂乳为基础培养基,通过正交试验方法优化得出ST4最佳增殖培养基配方为酵母粉0.5%、番茄汁7.5%、麦芽汁7.5%、乳清粉7.5%,活菌数达到1.72×109cfu/mL。正交试验优化所得ST4的最佳保护剂配方为海藻糖20%、乳糖2.5%、谷氨酸钠5%、甘油0.5%,经冷冻干燥后其活菌数达到1.56×1011cfu/g。     

基于蔗糖代谢途径分析保加利亚乳杆菌的后酸化性能

弱后酸化能力是乳酸菌作为发酵剂使用时的重要特性。为研究保加利亚乳杆菌蔗糖代谢途径对发酵乳后酸化起到的作用，该研究比较了五株保加利亚乳杆菌的后酸化性能，分析了保加利亚乳杆菌的生长情况、糖代谢和产酸能力，探究了蔗糖代谢产酸相关基因的差异表达及关键酶活性，并研究了外源添加蔗糖代谢关键酶前后保加利亚乳杆菌的生长情况。结果表明，Lb. 1后酸化能力最弱，在以蔗糖为碳源的培养基中生长缓慢，KLDS 1.0207后酸化能力最强。发酵24 h后，二者蔗糖转化率分别为5.85%和85.39%，乳酸含量分别为1.13 g/L和11.81 g/L。在含双糖（乳糖:蔗糖=1:1.5）的MRS培养基中生长时，KLDS 1.0207蔗糖代谢途径中基因sacA、pgi、gap、pgk、ldh表达量极显著高于Lb. 1（P<0.01），KLDS 1.0207蔗糖酶活性显著高于Lb. 1（P<0.05），达到1.31 U/mg。在Lb. 1的蔗糖培养基中补充蔗糖酶后，OD600 nm增至原来的5倍。因此，蔗糖酶活性对保加利亚乳杆菌代谢蔗糖至关重要，编码蔗糖酶的sacA基因表达下调显著减弱蔗糖酶活性，菌株后酸化能力明显下降。