气囊在细菌中的生理功能、生物合成及应用研究进展

气囊是在水生细菌中广泛存在的一种具有刚性中空蛋白结构的特殊细胞器,不仅为水生细菌提供浮力,还对其在不利环境或应激条件下的生存至关重要。近期研究发现在其他非水生细菌如沙雷氏菌和链霉菌中也存在气囊结构,而且表现出不同的生理功能。来源于不同种属细菌的气囊生物合成基因簇具有各自鲜明的特征,其生物合成和调控机制也有所不同。本综述将介绍和总结不同细菌中气囊的基本生理功能和生物合成及调控机制,以及气囊的生物技术应用,并对气囊在链霉菌中的生物合成研究以及人工重组气囊的潜在应用进行展望。     

人工蓝藻结皮对沙区表层土壤酶活性及其恢复速率的影响

土壤酶是土壤养分循环的重要参与者,也是反映土壤功能的重要指标之一。人工生物土壤结皮(Biological Soil Crusts, BSCs)是近年来新型的固沙技术之一,能够显著促进荒漠生态系统功能的恢复。然而,目前关于人工BSCs如何影响沙区土壤酶活性及恢复速率的研究仍鲜见报道。通过对腾格里沙漠东南缘人工培育以及自然发育的蓝藻结皮和流沙表层(0—2 cm)土壤酶活性(蔗糖酶、纤维素酶、淀粉酶、过氧化氢酶)以及BSCs的基本特征(叶绿素a和胞外多糖)进行测定,探讨了人工蓝藻结皮表层土壤酶活性的变化与恢复速率以及土壤酶活性与人工蓝藻结皮基本特征的关系。结果表明：人工蓝藻结皮表层土壤蔗糖酶(13.03—20.51 mg d^-1 g^-1)、纤维素酶(45.60—47.20 mg d^-1 g^-1)、过氧化氢酶(12.43—23.31μmol d^-1 g^-1)和淀粉酶活性(91.04—153.93 mg d^-1 g^-1),...     

朱鹮肠道微生物多样性与产酶活性

【背景】朱鹮是我国国家一级保护动物,属于世界上最濒危的鸟类之一。对朱鹮肠道微生物的多样性和产胞外酶活性进行分析,可为朱鹮种群数量恢复提供思路。【目的】了解朱鹮肠道微生物的多样性,测定其产胞外酶活性。【方法】采用纯培养的方法获得朱鹮肠道微生物,通过革兰氏染色和生理生化鉴定,结合16S rRNA基因扩增和序列分析对细菌进行鉴定。使用水解圈法筛选产淀粉酶、蛋白酶、纤维素酶、脂肪酶的菌株。【结果】从人工喂养的朱鹮新鲜粪便中共分离到296株细菌,共计2个门11个属。变形菌门(Proteobacteria) 236株,占分离总数的79.73%,分别为：埃希氏菌属(Escherichia) 137株,占分离总数的46.28%;哈夫尼亚菌属(Hafnia) 39株,占分离总数的13.18%;变形菌属(Proteus) 28株,占分离总数的9.46%;柠檬酸杆菌属(Citrobacter) 23株,占分离总数的7.77%;气单胞菌属(Aeromonas) 6株,占分离总数的2.03%;肠杆菌属(Enterobacter) 1株,占分离总数的0.34%;志贺菌属(Shigella) 1株,占分离总数的0.34%;克雷伯菌属(Klebsiella) 1株,占分离总数的0.34%。厚壁菌门(Firmicutes) 60株,占分离总数的20.27%,分别为：肠球菌属(Enterococcus) 33株,占分离总数的11.15%;库特氏菌属(Kurthia) 14株,占分离总数的4.73%;芽孢杆菌属(Bacillus) 13株,占分离总数的4.39%。优势菌群为变形菌门(Proteobacteria)中的埃希氏菌属(Escherichia),占细菌总数的46.28%。经过生理生化鉴定,每个菌株生理生化鉴定出的种属与各自的16S rRNA基因鉴定出的种属相一致。产酶活力分析结果显示有238株产蛋白酶、25株产脂肪酶、24株产淀粉酶、15株产纤维素酶,分别占分离总数的80.41%、8.45%、8.11%和5.07%。【结论】朱鹮肠道微生物分离出的细菌可分为2门11属,优势菌群为变形菌门(Proteobacteria)中的埃希氏菌属(Escherichia),占细菌总数的46.28%;产酶活性分析显示,80.41%的菌株具有产蛋白酶能力。     

五种矿质元素对草菇菌丝生长和活性氧清除能力的影响

【背景】菌种退化是草菇生产中面临的难题,探究一种简单高效的复壮方法是草菇产业亟须解决的问题。【目的】探讨外源添加5种矿质元素对草菇退化菌株的菌丝性状和活性氧(reactive oxygen species, ROS)清除能力的影响。【方法】筛选出5种矿质元素的最佳浓度并添加于PDA培养基中,测定草菇菌落形态、菌丝生长速度、菌丝生物量、ROS含量、抗氧化酶活性及抗氧化物质含量。【结果】MnSO4、Na2SeO3、CaSO4、FeSO4可有效提高草菇退化菌株D1和D2的菌丝生长速度和生物量;降低O2·-、H2O2等ROS的含量,并增加还原型谷胱甘肽(glutathione, GSH)、氧化型谷胱甘肽(glutathionedisulfide,GSSG)的含量及超氧化物歧化酶(superoxidedismutase,SOD)、过氧化氢酶(catalase, CAT)、谷胱甘肽...     

配合饲料和鲢鱼肉对大鲵生长的比较试验

为探讨人工配合饲料和鲢鱼肉对大鲵生长、消化和抗氧化能力的影响,试验选取初始体重为(20.99±0.15) g的大鲵幼体48尾,随机分成2组,每组3个重复,每个重复8尾,分别投喂人工配合饲料(粗蛋白55.67%,粗脂肪6.83%)和鲢鱼肉(粗蛋白18.03%,粗脂肪4.11%)共92d。结果显示:(1)饲料组大鲵的增重率(WGR)、特定生长率(SGR)、蛋白质沉积率(PRR)和肌肉蛋白质合成能力均显著高于鱼肉组,饲料系数(FCR)和存活率(SR)在两个试验组之间无显著差异(P>0.05)。(2)在大鲵摄食人工配合饲料后,全鲵粗蛋白、皮肤胶原蛋白及粗灰分含量均显著高于鱼肉组(P<0.05),而全鲵水分、粗脂肪与肌肉的粗脂肪和粗灰分含量显著低于鱼肉组(P<0.05)。(3)饲喂鲢鱼肉的大鲵胃蛋白酶活性显著高于饲料组(P<0.05),而两个试验组的胃H⁺-K⁺-ATP酶和肠道消化酶活性均无显著差异(P>0.05)。(4)饲喂人工配合饲料的大鲵肝脏总抗氧化能力(TAOC)和超氧化物歧化酶(SOD)活性显著高于鱼肉组(P&l...     

An Overview of a Wide Range of Functions of ZnT and Zip Zinc Transporters in the Secretory Pathway

Zinc plays essential roles in the early secretory pathway for a number of secretory, membrane-bound, and endosome/lysosome-resident enzymes. It enables the enzymes to fold properly and become functional, by binding as a structural or catalytic component. Moreover, zinc secreted from the secretory vesicles/granules into the extracellular space has a pivotal role as a signaling molecule for various physiological functions. Zinc transporters of the Slc30a/ZnT and Slc39a/Zip families play crucial roles in these functions, mediating zinc influx to and efflux from the lumen of the secretory pathway, constitutively or in a cell-specific manner. This paper reviews current knowledge of the ways these two zinc transporters perform these tasks by manipulating zinc homeostasis in the secretory pathway. Recent questions concerning zinc released into the cytoplasm from the secretory pathway, which then functions as an intracellular signaling molecule, are also briefly reviewed, emphasizing zinc transporter functions.     

Crystal structure of AdoMet radical enzyme 7‐carboxy‐7‐deazaguanine synthase from Escherichia coli suggests how modifications near [4Fe–4S] cluster engender flavodoxin specificity

7‐Carboxy‐7‐deazaguanine synthase, QueE, catalyzes the radical mediated ring contraction of 6‐carboxy‐5,6,7,8‐tetrahydropterin, forming the characteristic pyrrolopyrimidine core of all 7‐deazaguanine natural products. QueE is a member of the S‐adenosyl‐L‐methionine (AdoMet) radical enzyme superfamily, which harnesses the reactivity of radical intermediates to perform challenging chemical reactions. Members of the AdoMet radical enzyme superfamily utilize a canonical binding motif, a CX₃CX?C motif, to bind a [4Fe‐4S] cluster, and a partial (β/α)₆ TIM barrel fold for the arrangement of AdoMet and substrates for catalysis. Although variations to both the cluster‐binding motif and the core fold have been observed, visualization of drastic variations in the structure of QueE from Burkholderia multivorans called into question whether a re‐haul of the defining characteristics of this superfamily was in order. Surprisingly, the structure of QueE from Bacillus subtilis revealed an architecture more reminiscent of the classical AdoMet radical enzyme. With these two QueE structures revealing varying degrees of alterations to the classical AdoMet fold, a new question arises: what is the purpose of these alterations? Here, we present the structure of a third QueE enzyme from Escherichia coli, which establishes the middle range of the spectrum of variation observed in these homologs. With these three homologs, we compare and contrast the structural architecture and make hypotheses about the role of these structural variations in binding and recognizing the biological reductant, flavodoxin. Broader impact statement: We know more about how enzymes are tailored for catalytic activity than about how enzymes are tailored to react with a physiological reductant. Here, we consider structural differences between three 7‐carboxy‐7‐deazaguanine synthases and how these differences may be related to the interaction between these enzymes and their biological reductant, flavodoxin.     

Biocleaning on Cultural Heritage: new frontiers of microbial biotechnologies

Over the last two decades, the biotechnologies applied to Cultural heritage (CH) have become a successful novel alternative to the traditional approaches in the CH conservation and preservation. From these new perspectives, microorganisms and their metabolisms can be used for the safeguarding of artworks. Biocleaning is a field with a growing interest, based on eco-friendly processes and safe procedures, where biological reactions occurring in natural habitats are optimized in artificial conditions with the aim of CH conservation. This represents a new tool and opportunity for the development and improvement of the sector, with a great advantage for the CH conservation-restoration, in terms of safety, effectiveness, costs and environmental sustainability. This review focuses on the use of microbes and enzymes involved in biocleaning of CH artworks. The aim is to provide a comprehensive, critical and chronological view of the scientific works published until now where ‘virtuous’ microorganisms are applied on different CH materials, pointing out strength and drawback of the biocleaning treatments.     

Strategies for the production of cell wall‐deconstructing enzymes in lignocellulosic biomass and their utilization for biofuel production

Microbial cell wall‐deconstructing enzymes are widely used in the food, wine, pulp and paper, textile, and detergent industries and will be heavily utilized by cellulosic biorefineries in the production of fuels and chemicals. Due to their ability to use freely available solar energy, genetically engineered bioenergy crops provide an attractive alternative to microbial bioreactors for the production of cell wall‐deconstructing enzymes. This review article summarizes the efforts made within the last decade on the production of cell wall‐deconstructing enzymes in planta for use in the deconstruction of lignocellulosic biomass. A number of strategies have been employed to increase enzyme yields and limit negative impacts on plant growth and development including targeting heterologous enzymes into specific subcellular compartments using signal peptides, using tissue‐specific or inducible promoters to limit the expression of enzymes to certain portions of the plant or certain times, and fusion of amplification sequences upstream of the coding region to enhance expression. We also summarize methods that have been used to access and maintain activity of plant‐generated enzymes when used in conjunction with thermochemical pretreatments for the production of lignocellulosic biofuels.