适冷微生物及其适冷机制研究进展

地球上许多生境为永久低温或季节性低温环境,适冷微生物在自然界中广泛存在。适冷微生物在环境净化、饲料、食品、奶制品、化妆品、皮革加工、洗涤等行业中具有广泛的应用前景。对适冷微生物的多样性、适冷的分子基础和适冷代谢机制进行了综述。     

低温微生物及其酶类的研究概况

广泛分布在地球寒冷生境 ,如南北两极、高山、深海以及冰川中的低温微生物 ,不但为研究低温生态系统、生命起源与进化以及生物适冷机制提供了丰富的材料 ,同时在生物工程方面也具有潜在的巨大开发价值。国内外越来越多的科研人员对低温微生物及其产物的研究表现出了浓厚的兴趣。关于细胞膜和低温酶的研究 ,是目前微生物适冷机制研究中的 2个热点。就低温微生物的研究现状和适冷机制以及低温酶类的研究进行了综述。     

微生物适冷酶及其应用研究新进展

生物圈的80%是由低温环境构成,大约90%的海水平均温度为5℃或者更低,这里孕育着极为丰富的微生物。根据微生物对环境温度的耐受性和其生长温度上限/下限的不同,将微生物分为嗜冷微生物和适冷微生物两大类。由这些微生物产生的适冷酶在低温下具有较高的催化效率和特异性,因而在生物技术领域具有巨大的应用潜力和开发价值。总结了近5年适冷酶新酶的筛选、发现及适冷酶稳定性的改造和低温表达系统等方面的最新研究进展,并对该领域的研究方向进行了展望。     

常温土壤中耐冷菌的分离、鉴定及产酶分析

目的:从常温土壤中筛选冷适应微生物,并进行初步鉴定和产低温酶分析。方法:采集吉首大学校园内土壤样品,通过低温富集培养筛选冷适应微生物;通过形态观察、生理生化特性检测和基于16S rRNA基因序列的系统发育分析,对分离的菌株进行初步鉴定;利用平板筛选法检测其产低温酶特性。结果:分离获得6株耐冷细菌JSBP-1～JSBP-6,初步鉴定其分属假单胞菌属(Pseudomonas)、紫色杆菌属(Janthinobacterium)和节杆菌属(Arthrobacter);在5℃和15℃培养条件下,菌株JSBP-1产蛋白酶能力较强,JSBP-2和JSBP-6产淀粉酶能力较强,JSBP-5仅在5℃条件下有较强的产脂肪酶特性。结论:常温土壤中存在一定数量的冷适应微生物,其中假单胞菌是其优势菌群之一。这类适冷微生物菌群具有潜在的生产低温酶能力。     

嗜冷酶及其工业应用

生物圈中超过 80 %的地方温度低于5℃ ,在生态学上低温环境的影响范围更广。生活在这些低温环境中的嗜冷菌对低温有特殊的适应性。相对于嗜温菌而言 ,嗜冷微生物在低温条件下调节其膜流动性和膜通透性的能力更强 ,含有更多的不饱和脂肪酸和短链脂肪酸 ,还可通过合成冷激蛋白来响应冷激作用。嗜冷菌也可通过基因调控和自身的抗生素作用来适应低温。近来有研究报道 ,南北极严寒地区的海洋鱼类依靠血液中的抗冻蛋白来适应低温。生物体内的大多数生化反应都是由酶催化而成的 ,所以考虑嗜冷微生物的适应机制时首先应研究嗜冷酶的适冷机制。1 .嗜…     

低温微生物适应低温的分子机制

根据Ｍｏｒｉｔａ和Ｒｕｓｓｅｌ的定义 ,低温微生物可分为两类 :一类是其最高生长温度低于2 0℃的微生物称嗜冷菌 (ｐｓｙｃｈｒｏｐｈｉｌｅｓ) ,另一类是指在 0～ 40℃可以生长的微生物称耐冷菌 (ｐｓｙｃｈｒｏｔｒｏｐｈｓ)。这些微生物体内的温度与环境温度很接近。尽管低温对生化反应有着强烈的负效应 ,但低温微生物在低温下却能成功地生长与繁殖 ,它们因此产生了各种各样的适应机制 ,主要是细胞膜、蛋白质、酶分子水平上发生了精细的组成和结构的变化[1] ,因此弥补了低温对生长的有害影响。催化生物体内所有生化反应的酶是生物体适…     

低温细菌与古菌的生物多样性及其冷适应机制

低温细菌与古菌广泛分布于地球的低温环境,包括南极、北极及高山地带的冻土、低温土壤和荒漠、冰川、湖泊、海冰,以及深海、冰洞和大气平流层等.栖息在这些低温环境中的细菌与古菌具有丰富的多样性,主要为α,p和γ-Proteobacteria分支、CFB类群分支和革兰氏阳性细菌分支等.由于低温环境中的微生物流动性低,因而是研究微生物地理学理想的生态系统,有助于理解地球微生物的多样性、分布规律乃至形成机制.由于长期生活在冰冻环境中,低温细菌与古菌形成了多种适应低温环境的生理机制,如它们通过细胞膜脂类的组成来调节膜的流动性以维持正常的细胞生理功能;利用相容性溶质、抗冻蛋白、冰核蛋白及抗冰核形成蛋白等实现低温保护作用;产生冷激蛋白、冷适应蛋白和DEAD-box RNA解旋酶保持低温下RNA的正确折叠、蛋白质翻译等重要的生命活动;另外还产生低温酶,提高能量产生和储存效率等以适应低温环境.随着DNA序列分析技术的飞速发展,各类组学方法也用于揭示微生物全局性的冷适应机制.     

微生物产生的冷休克蛋白研究进展

冷休克蛋白(cold shock protein,Csp)首先在大肠杆菌中发现,它与微生物对冷环境的适应及多种细胞功能有关。冷休克蛋白基因是一段编码70个左右氨基酸的DNA序列,在这段序列中有5′非翻译区(5′UTR)、冷盒及下游盒等特征。冷休克蛋白作为DNA或RNA结合蛋白在基因表达调控过程中起重要作用。冷休克蛋白在转录、mRNA稳定性及翻译等几个水平上被严格调控。     

微生物对低温极端环境适应性的研究进展

嗜冷微生物是地球寒冷环境中最主要的生物类群,并且是驱动全球生物地球化学循环的关键环节。嗜冷微生物在适应策略上显示出应对多种极端环境因素的巨大潜力,研究其适应和进化机制有助于更好地理解微生物与环境之间相互作用过程,并有效利用极端环境微生物资源。近年来,随着分子生物学和基因组学技术的高速发展,对微生物适应寒冷环境的机制及嗜冷微生物在指示气候变化和工农业应用方面均有一系列的突破。在此,本文将从基因组的GC含量、蛋白质稳定性、转录翻译调控、细胞膜流动性、渗透压调节、抗氧化损失和基因组适应性进化等方面总结当前在微生物适应低温环境机制上所取得的进展,并展望低温环境微生物在指示气候变化和工农业应用中的前景。     

冷激蛋白在极地微生物冷适应中的作用

冷激蛋白对极地微生物的低温生境适应性起着重要作用。该介绍了冷激蛋白的广泛性、冷激特性与功能,以及冷诱导调控研究的最新进展。     

蛋白质的变化与植物抗寒性的关系研究进展

蛋白质的变化在植物抗寒生理研究中一直被广泛关注。低温胁迫期间在蛋白质含量变化的同时,还可能发生质的变化,合成新的蛋白质——低温诱导蛋白。综述了低温胁迫期间植物体内蛋白质的变化,重点阐述了抗冻蛋白、脱水蛋白和热激蛋白等3种低温诱导蛋白的特性及其与植物抗寒性的关系,并对该领域今后的研究做了展望,为进一步阐明植物抗寒的分子机制、提高植物的抗寒力提供了新的思路。     

低温诱导蛋白及其与植物的耐寒性研究进展

低温诱导蛋白是植物在温度逆境条件下诱导产生的一系列蛋白,以抗冻蛋白、脱水蛋白、热激蛋白和热稳定蛋白较多,而且低温诱导蛋白质一旦在体内形成,植物体就会尽快地适应外界环境,表现出较强的抗逆性.本文对几种主要的低温诱导蛋白——抗冻蛋白、脱水蛋白、热激蛋白和热稳定蛋白的特性及其与植物耐寒性的关系研究进行综述,以期为进一步阐明植物耐寒的分子机制以及提高植物耐寒力研究提供新的思路.     

多不饱和脂肪酸对微生物低温适应性的影响

地球生物圈75%以上的环境温度常年低于5℃,在这种低温环境中栖息着多种适应低温的微生物。在长期进化过程中低温微生物从细胞到分子水平形成一套独特的低温环境适应机制,而通过增加细胞膜膜脂中多不饱和脂肪酸含量来维持低温条件下最佳的细胞膜流动性是其中的一种。从多不饱和脂肪酸对微生物低温生长、细胞膜流动性细胞膜蛋白的组成和表达水平的影响来探讨多不饱和脂肪酸与微生物低温适应性的关系,总结多不饱和脂肪酸低温合成调节机制的研究进展,为相关的基础和应用开发研究提供参考。     

昆虫冷驯化机制研究进展

昆虫耐寒性强弱决定其种群的发生、扩散和分布,因此低温胁迫下昆虫的抗寒对策成为近期研究的热点领域。冷驯化作为一种非常有效的耐寒策略,可显著增强昆虫的耐寒性。本文论述了冷驯化的2种基本形式:快速冷驯化和长时冷驯化,明确了二者在提升昆虫耐寒性中的作用;并从宏观到微观的角度概述了冷驯化的作用机制,如组织和细胞水平的特异性,低分子量抗冻保护剂的产生,热休克蛋白的表达及功能,以及阻止细胞程序性死亡的潜在机理等;讨论了不同研究方法所引起的结果差异性,并强调了冷驯化作用机制的整体效益和综合效益。最后通过分析2种冷驯化形式的联系与区别,以期较为全面地阐明昆虫冷驯化的潜在机制。     

抗寒类蛋白与冷驯化诱发昆虫耐寒性研究进展

昆虫是变温动物,温度对其生长发育、基本行为及进化途径都会产生很大的影响,种群的繁衍面临如何安全度过漫长而寒冷的冬季的挑战。通过长时间的进化,昆虫获得一系列完整的耐寒策略。绝大多数的昆虫都是耐寒昆虫,在陆地寒冷温度刺激下,昆虫受抗寒基因的调控,体内产生大量抗寒物质,如海藻糖、甘油、山梨醇、抗冻蛋白、热激蛋白等,提高昆虫的耐寒能力,使其得以在低温寒冷的条件下成功越冬。同样,经过冷驯化后的昆虫能显著提高昆虫的耐寒力。近年来,关于昆虫耐寒性、抗寒类蛋白的研究不断开展,研究内容涉及昆虫的耐寒性、抗寒基因HSPs和AFPs的调控、冷驯化诱导抗寒等方面。本文综述了昆虫耐寒性、主要耐寒策略及冷驯化诱发昆虫耐寒性增强等研究内容。有助于全面认识昆虫耐寒性及其作用机制,为天敌昆虫低温储存和提高生物防治等应用打下坚实的基础。     

Cloning of Fish Enzymes and Other Fish Protein Genes

ABSTRACT:?

Fish metabolism needs special enzymes that have maximum activity at very different conditions than their mammalian counterparts. Due to the differences in activity, these enzymes, especially cold-adapted proteases, could be used advantageously for the production of some foods. In addition to the enzymes, this review describes some other unique fish polypeptides such as antifreeze proteins, fluorescent proteins, antitumor peptides, antibiotics, and hormones, that have already been cloned and used in food processing, genetic engineering, medicine, and aquaculture. Recombinant DNA technology, which allows these biological molecules to be cloned and overexpressed in microorganisms is also described, highlighting innovative applications. The expected impact of cloning fish proteins in different fields of technology is discussed.     

PROTEOMICS ANALYSIS OF OVEREXPRESSED PLASMA PROTEINS IN RESPONSE TO COLD ACCLIMATION IN Ostrinia furnacalis

Many insects in temperate regions overwinter in diapause. In these insects, one of the metabolic adaptations to cold stress is the synthesis of responsive proteins. Using proteomic analysis, an investigation aimed to a better understanding of the molecular adaptation mechanisms to cold stress was carried out in Ostrinia furnacalis larva. Proteins were extracted from the larval hemolymph collected from both control and overwintering larva. By polyethylene glycol precipitation, approximately 560 protein spots were separated and visualized on two‐dimensional (2D) gels after silver staining. Eighteen protein spots were found to be upregulated in overwinter larval plasma in different patterns. As an initial work, 13 of these proteins were identified using MALDI TOF/TOF MS. The differentially overexpressed proteins include heat shock 70 kDa cognate protein, small heat shock protein (sHSP), putative aliphatic nitrilase, arginine kinase, phosphoglyceromutase, triosephosphateisomerase, and glutathione transferase. Alterations in the levels of these proteins were further confirmed by qPCR. This study is the first analysis of differentially expressed plasma proteins in O. furnacalis diapause larvae under extremely low temperature conditions and gives new insights into the acclimation mechanisms responsive to cold stress. Our results also support the idea that energy metabolism, alanine and proline metabolism, and antioxidative reaction act in the cold acclimation of O. furnacalis diapause larvae.     

The effects of ionic strength on protein stability: the cold shock protein family

Continuum electrostatic models are used to examine in detail the mechanism of protein stabilization and destabilization due to salt near physiological concentrations. Three wild-type cold shock proteins taken from mesophilic, thermophilic, and hyperthermophilic bacteria are studied using these methods. The model is validated by comparison with experimental data collected for these proteins. In addition, a number of single point mutants and three designed sequences are examined. The results from this study demonstrate that the sensitivity of protein stability toward salt is correlated with thermostability in the cold shock protein family. The calculations indicate that the mesophile is stabilized by the presence of salt while the thermophile and hyperthermophile are destabilized. A decomposition of the salt influence at a residue level permits identification of regions of the protein sequences that contribute toward the observed salt-dependent stability. This model is used to rationalize the effect of various point mutations with regard to sensitivity toward salt. Finally, it is demonstrated that designed cold shock protein variants exhibit electrostatic properties similar to the natural thermophilic and hyperthermophilic proteins.     

Role of HSF activation for resistance to heat, cold and high-temperature knock-down

Regulation of heat shock proteins (Hsps) by the heat shock factor (HSF) and the importance of these proteins for resistance to heat stress is well documented. Less characterized is the importance of Hsps for cold stress resistance although Hsp70 is known to be induced following long-term cold exposure in Drosophila melanogaster. In this study, a temperature-sensitive HSF mutant line was used to investigate the role of HSF activation following heat hardening, rapid cold hardening (RCH) and long-term cold acclimation (LTCA) on heat and cold resistance, and this was correlated with Hsp70 expression. In addition, the effect of HSF activation on high-temperature knock-down resistance was evaluated. We found a significantly decreased HSF activation in the mutant line as compared to a corresponding control line following heat hardening, and this was correlated with decreased heat resistance of the mutant line. However, we did not find this difference in HSF activity to be important for resistance to cold stress or high-temperature knock-down. The findings indicate that induction of stress genes regulated by HSF, such as Hsps, although occurring following LTCA, are not of major importance for cold stress resistance and neither for RCH nor high-temperature knock-down resistance in D. melanogaster.     

Adaptation to low temperature and regulation of gene expression in antarctic psychrotrophic bacteria

Exposure to extremes of temperatures cause stresses which are sometimes lethal to living cells. Microorganisms in nature, however, are extremely diverse and some of them can live happily in the freezing cold of Antarctica. Among the cold adapted psychrotrophs and psychrophiles, the psychrotrophic bacteria are the predominant forms in the continental Antarctica. In spite of living in permanently cold area, the antarctic bacteria exhibit, similar to mesophiles, ‘cold-shock’ response albeit at a much lower temperatures, e.g., at 0–5°C. However, because of permanently cold condition and the long isolation of the continent, the microorganisms have acquired new adaptive features in the membranes, enzymes and macromolecular synthesis. Only recently these adaptive modifications are coming into light due to the efforts of various laboratories around the world. However, a lot more is known about adaptive response to low temperature in mesophilic bacteria than in antarctic bacteria. Combined knowledge from the two systems is providing useful clues to the understanding of basic biology of low temperature growing organisms. This article will provide an overview of this area of research with a special reference to sensing of temperature and regulation of gene expression at lower temperature.