High Produciton and Application of a Novel Ice-nucleating Matter from Microorganisms

新型微生物冰核物质的高产及应用

• 批准号: 09555258
• 负责人: OBATA Hitoshi
• 金额: $ 3.14万
• 依托单位: KANSAI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09555258/
• 关键词: Xanthomonas; ice-uncleating matter; xanthan gum; Pseudomonas; extracellular ice-nucleating matter; freeze-concentration; 氷核タンパク質; クラスA,B,C; 氷核活性細菌; X-compestris; 人工降雪剤; 食品素材改良剤

Xanthomonas campestric pv. translucens IF013599 could produce xanthan gum (18.5 mg/100 mg, lactose) with lactose as the growth substrate in spite of a low level of beta-galactosidsse. This productivity contspondsd to one-fifth of that with glucose. This strain could also produce ice-nucleating matter having an ice-nucleating temperature, T50, of -2.8* with xanthan gum in the culture broth. We found that this strain produced both matenals in the whey medium from which the insoluble components had been removed The production of xanthan with ice-nucleating material reached a maximum after cultivation for 168 h uuder optimum conditions. Furthennore, the xanthan obtained had a low viscosity because of its variant structure revealed, by the TLC and HPLC analyses, to be lacking pyruvic acid.Also, the extracellular ice-nucleating matter produced by the ice-nucleating bacterium, Pseudomonos fluorescens KUIN-1, was punfiedto an apparent homogeneity. The shape of this EIM was spherical with the d … More iameter of 10-100 nm without a vesicle. The ice-nucleating activity and the stability of this EIM were similar to that of cell in this strain. It had become apparent that this ElM had class A (wanner at -5.0ﾟC), B(-6--7ﾟC) and C(coldur at -7.5ﾟC) stnictures constituting different components like that of the ElM from Erwinia uredovora KUIN-3. The components of the ElM were protein (56%), lipid (15%), saccharide (10%) and polyamine (19%). Based on the SDS-PAGE analysis with Western-blotting, the main band, that is, for ice-nucleating protein (INP), was 110,000 Da, and minor bands were also attributed to the possible INP-degrading proteins. The fatty acid components of the lipids were the same as those of the inner and outer membranes. The saccharied was composed of glucose, galactosamine, rhamnose, etc. Cachverine, spermidine and putrescine were present as the components of the EIM.This EIM had the distinct ability of freeze-concentrating wine and vinegar. This is the first report for the purification of the EIM that is produced by an ice-nucleating bacterium belonging to the genus Pseudomonas and for the application of the ElM in freeze-concentration of foods. Less

葡萄黄单胞菌。translucens IF013599在β -半乳糖苷含量较低的情况下，仍能以乳糖为底物生产黄原胶（18.5 mg/100 mg，乳糖）。这种生产力相当于葡萄糖的五分之一。该菌株在黄原胶的培养液中也能产生冰核物质，冰核温度T50为-2.8*。我们发现，该菌株在去除不溶性成分的乳清培养基中产生了这两种物质，冰核材料在最佳条件下培养168 h后黄原胶的产量达到最大。此外，通过薄层色谱和高效液相色谱分析，得到的黄原胶具有低粘度，因为其结构不同，缺乏丙酮酸。此外，由冰核细菌产生的细胞外冰核物质，荧光假单胞菌KUIN-1，被刺激到明显的均匀性。该EIM的形状为球形，直径为10 ~ 100 nm，无囊泡。该EIM的冰核活性和稳定性与该菌株的细胞相似。很明显，这种榆树具有A级（-5.0 C）， B级（-6- 7 C）和C级（-7.5 C）结构，构成不同的成分，与Erwinia uredoora KUIN-3中的榆树相似。榆木的主要成分为蛋白质（56%）、脂质（15%）、糖类（10%）和多胺（19%）。Western-blotting的SDS-PAGE分析显示，冰核蛋白（INP）的主要条带为110,000 Da，次要条带也归属于可能的INP降解蛋白。脂质脂肪酸组成与内、外膜相同。该糖剂由葡萄糖、半乳糖胺、鼠李糖等组成。恶毒碱、亚精胺和腐胺作为EIM的组成部分存在。该EIM具有明显的冷冻浓缩酒和醋的能力。这是首次报道由假单胞菌属的冰核细菌产生的EIM的纯化以及ElM在食品冷冻浓缩中的应用。少

项目成果

Hidehisa Kawahara: "Purification and Characterization of the Extracellular Ice-nucleating Matter from the Ice-nucleating Bacterium, Pseudomonas fluorescens KUIN-1" Applied Microbiology and Biotechnology. (in press). (1999)

Hidehisa Kawahara：“冰核细菌荧光假单胞菌 KUIN-1 细胞外冰核物质的纯化和表征”应用微生物学和生物技术。

Hidehisa Kawahara and Hitoshi Obata: "Production of Xanthan Gum and Ice-nucleating Material from Whey by Xanthomonas campestris pv. translucens" Applied Microbiology and Biotechnology. 49. 353-358 (1998)

Hidehisa Kawahara 和 Hitoshi Obata：“利用 Xanthomonas Campestris pv. translucens 从乳清中生产黄原胶和冰成核材料”应用微生物学和生物技术。

Hidehisa Kawahara and Hitoshi Obata: "Production of Xanthan Gum and Ice-nucleating Material from Whey by Xanthomonas campestris pv.translucens" Applied Microbiology and Biotechnology. 49. 353-358 (1998)

Hidehisa Kawahara 和 Hitoshi Obata：“利用 Xanthomonas Campestris pv.translucens 从乳清中生产黄原胶和冰成核材料”应用微生物学和生物技术。

Hidehisa Kawahara, Mariko Matsushita, Kazuhiro Yamade and Hitoshi Obata: "Purification and Characterization of the Extracellular Ice-nucleating Matter from the Ice-nucleating Bacterium, Pseudomonas fluorescens KUIN-1" Applied Microbiology and Biotechnolog

Hidehisa Kawahara、Mariko Matsushita、Kazuhiro Yamade 和 Hitoshi Obata：“冰核细菌荧光假单胞菌 KUIN-1 细胞外冰核物质的纯化和表征”应用微生物学和生物技术

Hidehisa Kawahara: "Production of Xanthan Gum and Ice-nucleating Materials from Whey by Xanthomonas campestris pv.translucens" Applied Microbiology and Biotechnology. 49. 353-358 (1998)

Hidehisa Kawahara：“利用 Xanthomonas Campestris pv.translucens 从乳清中生产黄原胶和冰成核材料”应用微生物学和生物技术。