细菌纤维素生产菌株的分离和菌种初步鉴定

从长膜的醋醅中分离出一株发酵生产细菌纤维素产量较高且稳定的醋酸菌M12。根据《常见细菌系统鉴定手册》和《伯杰氏细菌鉴定手册》第九版,对醋酸菌M12进行了形态和生理生化特征的分析、测定了G＋Cmol％含量,初步鉴定该菌为醋化醋杆菌木质亚种（Acetobacter xylinum subsp.xylinum,又称木醋杆菌）。     

日本中野醋店和东京大学确立了醋化醋杆菌基因自体克隆技术

日本中野醋店已确立了食醋商业化生产所用的醋化醋杆菌(醋酸菌)(Acetobacter acefi)的有用基因自体克隆的基础技术。利用这项成果,就可使由限制性酶切断的DNA片断连接到载体上使其克隆化,使从得到的许多克隆通过散弹枪法(shot gun)克隆找出目标基因的技术能在醋酸菌中进行,从而可加快醋酸菌有用基因的克隆化。该公司正利用基因重组技术从事这种菌的育种工作:能降低醋酸发酵的冷却成本的高温发酵醋酸菌,有利于业务用食醋的高酸度发酵醋酸菌,提高醋酸发酵生产速度的醋酸菌等。     

醋酸菌耐酸机理及其群体感应研究新进展

醋酸菌(acetic acid bacteria,AAB)是一类严格好氧的革兰氏阴性细菌,因其乙醇氧化生成醋酸能力强、高耐醋酸等特性而成为食醋发酵的主要工业菌种。醋酸菌的耐酸性对于高酸度食醋生产具有重要意义。随着醋酸菌的蛋白组学及基因组学研究的深入,其糖代谢、蛋白质代谢、脂代谢及应激响应等分子机制或过程也得到更多的阐释;葡糖醋杆菌中有关群体感应系统的研究报道则为从信号通路角度探索醋酸菌的耐酸机制提供了新的思路,进而对于高耐酸醋酸菌的选育以及醋酸发酵工艺的优化具重要的参考意义。本文在简介蛋白组、基因组研究的基础上,着重综述醋酸菌群体感应的研究进展。     

利用果渣产黄腐酸菌剂的筛选

以果渣发酵生产黄腐酸可实现农业有机固定废弃物的资源化利用。从腐植酸样品中分离获得4株耐高温菌,以果渣为基质发酵生产黄腐酸,通过测定黄腐酸产量和木质纤维素降解率,从中筛选出2株能够较好降解木质纤维素的黄腐酸生产菌株BFA02和BFA03,其黄腐酸产量分别为43.67、59.35 g/kg。经16S rDNA分类鉴定,初步确认BFA02为解淀粉芽孢杆菌,BFA03为地衣芽孢杆菌。选择季也蒙酵母、长枝木霉分别与BFA02、BFA03复配发酵生产黄腐酸。结果表明,复合菌剂发酵生产黄腐酸产量高于单菌发酵。其中,由BFA02、BFA03和长枝木霉组成的复合菌剂3实验组黄腐酸产量最高,达到103.19 g/kg,较BFA02、BFA03单菌发酵分别提高了136%和74%,其纤维素、半纤维素、木质素的降解率分别为46.25%、39.49%、37.5%,均高于单菌发酵。黄腐酸生产菌株与长枝木霉复配,能够有效促进木质纤维素降解率,提高黄腐酸产量,在果渣废弃物资源化利用领域有着极大的潜力。     

应用基因工程方法改良食醋

据日本《日经产业新闻》报道,日本最大的食醋工业公司中,醋店决定应用生物工程技术改良醋酸菌,并于本年七月向科学技术厅申请了对醋酸菌的重组基因实验。该企业改良醋酸菌的目的是:①使优良菌种具有在高温下的耐性,减少发酵过程中的冷却工艺,从而而节约能源;②能在酸度高的条件下发酵,或是加快发酵速度以提高生产;③改变食醋组成成分提高味感质量。     

高温嫌气纤维素分解细菌发酵纤维素的研究

我们研究了高温嫌气纤维素分解细菌的纯粹培养,分解纤维素的能力。在发酵过程中,培养15天时,纤维素分解串可达53％．延长培养时间,未见增加。培养20天产生的葡萄糖、醋酸、乳酸,达到最大量,分别为分斛纤维素的76％、12％、7％。该菌对碱性基质侑良好适应性,当培养液的pH增高到8.9时,纤维素分解率最高,生成的还原糖量亦较多。基质pH接近9.3时,二者均降低,但酸的生成量,仍然增加;此菌还能直接发酵加于培养基巾的葡萄糖,形成与发酵纤维紊相同的确机酸组分。并用分配层析法,对发酵液含的微盈有机酸作了测定,结果稳矗,可以清楚地分离混合存在的何机酸类。     

酵母菌和食用真菌混合固态发酵配合蛋白饲料的应用研究

双菌混合发酵是现代发酵技术发展趋势之一,高温酵母菌和担子茵混合固态发酵尝试了一种生产配合饲料的新技术,高温酵母菌生长速度快,适应性强,蛋白含量高;担子菌分解纤维素和半纤维素能力强,利用它们各自特点,经双菌协同混合发酵,不但提高了淀粉渣的蛋白质含量,而且也降低了纤维素和半纤维素含量,从而使无法直接利用的淀粉渣转化为优质配合蛋白饲料。      

应用基因工程方法改良食醋

据日本《日经产业新闻》报道,日本最大的食醋工业公司中(林土)醋店决定应用生物工程技术改良醋酸菌,并于本年七月向科学技术厅申请了对醋酸菌的重组基因实验。 该企业改良醋酸菌的目的是:①使优良菌种具有在高温下的耐性,减少发酵过程中的冷却工艺,从而节约能源;②能在酸度高的条件下发醋,或是加快发酵速度以提高生产;③改变食醋组成成份,提高味感质量。     

用生料制备功能性双歧醋生产工艺的实验室研究

目的将双歧杆菌、醋酸菌、酵母菌和粉碎的制醋原料及麸曲共同发酵,通过生料制醋的方法来制备功能性双歧醋。方法将粉碎的玉米与麸曲、酵母液、麸皮和水搅拌均匀,使其经过液态糖化和酒精发酵后,接入醋酸菌和双歧杆菌(二者比例为1∶1),同时加入辅料,进行醋酸发酵,当检测到醋酸酸度为5.0%~7.5%时,加入食盐终止发酵,经过过滤,除菌澄清得到功能性双歧醋。结果双歧醋的最终醋酸度为3.2%,外观红棕色,光泽度好,清澈透明,无沉淀和悬浮物。总菌数:醋酸菌为3.3×1011/m l,双歧杆菌为1.9×107/m l;活菌数:醋酸菌为1.7×1011/m l,双歧杆菌为6.8×106/m l;大肠菌群数3个/100 m l;致病菌:不得检出。结论双歧杆菌及其代谢物可以在双歧醋中存活,生料固态发酵制备双歧醋的方法可行。     

饲用微生物的分离鉴定及其对杏鲍菇菌糠发酵的效果

【目的】菌糠的营养素含量齐全,但纤维素含量过高是阻碍其饲料化利用的主要因素。故本研究筛选适合于发酵杏鲍菇菌糠的微生物菌株,以改善其饲用品质。【方法】首先,本研究采用纤维素-刚果红、苯胺蓝和MRS-Ca (De Man, Rogosa, Sharpe-Ca)筛选培养基,结合纤维素、木质素酶活力及抑菌活性的测定,从EM (effective microorganisms)原液发酵的杏鲍菇菌糠中分离筛选具有较强纤维素、木质素降解能力及抑菌能力的细菌/真菌。通过细菌16S rRNA和真菌18S rDNA基因序列分析确定菌株所属种属。其次,将筛选出的菌株菌液等体积混合制成复合菌剂用于固态发酵杏鲍菇菌糠。测定不同发酵时长菌糠营养成分含量以确定最佳发酵时间,并与相同工艺条件下EM原液发酵的杏鲍菇菌糠进行饲用品质比较。【结果】筛选并鉴定得到纤维素酶活性较高的特基拉芽孢杆菌(Bacillus tequilensis)菌株P11、发酵毕赤酵母(Pichia fermentans)菌株R8和马克斯克鲁维应变酵母(Kluyveromyces marxianus)菌株MU5;木质素酶活性较高的解淀粉芽孢杆菌(Bacillus amyloliquefaciens subsp.plantarum)菌株MU7;抑菌活性较高的类肠膜魏斯氏菌(Weissella paramesenteroides)菌株R4和乳酸片球菌(Pediococcus acidilactici)菌株R9。使用以上菌株复合发酵杏鲍菇菌糠7 d后,各项指标达到稳定。与EM原液发酵的杏鲍菇菌糠相比,复合菌剂发酵杏鲍菇菌糠的NDF和ADF分别显著降低了19.6%和21.44%(P0.05);CP (crude protein)、CA (crude ash)和EE (ether extract)含量分别显著提高了10.44%、5.26%和123.53%(P0.05)。【结论】本研究筛选得到的芽孢杆菌、酵母菌和乳酸菌优势菌株复合后用于发酵杏鲍菇菌糠可以很好地改善其饲用品质,效果优于生产中常用市售EM原液。     

细菌纤维素的研究进展

细菌纤维素是由醋酸杆菌属、根瘤菌属、土壤杆菌属、八叠球菌属等的某些细菌在一定条件下产生的,其中最有代表性的细菌是木醋杆菌。与传统植物纤维素相比,细菌纤维素具有很高的化学纯度。主要介绍细菌纤维素性质、生物合成的方法及其在食品工业、造纸工业和作为一种生物材料在医学工程等方面的应用。     

Effects of mixing conditions on the production of microbial cellulose byAcetobacter xylinum

Microbial cellulose has many potential applications due to its excellent physical properties. The production of cellulose fromAcetobacter xylinum in submerged culture is, however, beset with numerous problems. The most difficult one has been the appearance of negative mutants under shaking culture conditions, which is deficient of cellulose producing ability. Thus genetic instability ofAcetobacter xylinum under shaking culture condition made developing a stable mutant major research interest in recent years. To find a proper type of bioreactor for the production of microbial cellulose, several production systems were developed. Using a reactor system with planar type impeller with bottom sparging system, it was possible to produce 5 g/L microbial cellulose without generating cellulose minus mutants, which is comparable to that of static culture system.     

Structure of Acetobacter cellulose composites in the hydrated state

The structure of composites produced by the bacterium Acetobacter xylinus have been studied in their natural, hydrated, state. Small-angle X-ray diffraction and environmental scanning electron microscopy has shown that the ribbons have a width of 500 A and contain smaller semi-crystalline cellulose microfibrils with an essentially rectangular cross-section of approximately 10 x 160 A(2). Incubation of Acetobacter in xyloglucan or pectin results in no changes in the size of either the microfibrils or the ribbons. Changes in the cellulose crystals are seen upon dehydration of the material, resulting in either a reduction in crystal size or an increase in crystal disorder.     

Composites of bacterial cellulose and paper made with a rotating disk bioreactor

Suspended solids in the nutrient medium for Acetobacter xylinium in a rotating disk bioreactor become incorporated into the gelatinous mat of bacterial cellulose as it forms. Embedding fibers of ordinary cellulose creates composites with enhanced strength and the toughness of bacterial cellulose. Purified cellulose and elongated fibers from paper are incorporated differently than are spherical particles such as silica gel. About 90% of the final cellulose can come from scrap paper, and dried composite sheets were much stronger than plain bacterial cellulose per unit area.     

Characterization of genes in the cellulose-synthesizing operon (acs operon) of Acetobacter xylinum: implications for cellulose crystallization.

The synthesis of an extracellular ribbon of cellulose in the bacterium Acetobacter xylinum takes place from linearly arranged, membrane-localized, cellulose-synthesizing and extrusion complexes that direct the coupled steps of polymerization and crystallization. To identify the different components involved in this process, we isolated an Acetobacter cellulose-synthesizing (acs) operon from this bacterium. Analysis of DNA sequence shows the presence of three genes in the acs operon, in which the first gene (acsAB) codes for a polypeptide with a molecular mass of 168 kDa, which was identified as the cellulose synthase. A single base change in the previously reported DNA sequence of this gene, resulting in a frameshift and synthesis of a larger protein, is described in the present paper, along with the sequences of the other two genes (acsC and acsD). The requirement of the acs operon genes for cellulose production was determined using site-determined TnphoA/Kanr GenBlock insertion mutants. Mutant analysis showed that while the acsAB and acsC genes were essential for cellulose production in vivo, the acsD mutant produced reduced amounts of two cellulose allomorphs (cellulose I and cellulose II), suggesting that the acsD gene is involved in cellulose crystallization. The role of the acs operon genes in determining the linear array of intramembranous particles, which are believed to be sites of cellulose synthesis, was investigated for the different mutants; however, this arrangement was observed only in cells that actively produced cellulose microfibrils, suggesting that it may be influenced by the crystallization of the nascent glucan chains.     

Molecular interactions in bacterial cellulose composites studied by 1D FT-IR and dynamic 2D FT-IR spectroscopy

Specific strain-induced orientation and interactions in three Acetobacter cellulose composites: cellulose (C), cellulose/pectin (CP) and cellulose/xyloglucan (CXG) were characterized by FT-IR and dynamic 2D FT-IR spectroscopies. On the molecular level, the reorientation of the cellulose fibrils occurred in the direction of the applied mechanical strain. The cellulose-network reorientation depends on the composition of the matrix, including the water content, which lubricates the motion of macromolecules in the network. At the submolecular level, dynamic 2D FT-IR data suggested that there was no interaction between cellulose and pectin in CP and that they responded independently to a small amplitude strain, while in CXG, cellulose and xyloglucan were uniformly strained along the sample length.     

Inhibition of the synthesis of microbial cellulose by coumarin

Coumarin, a specific inhibitor of the biosynthesis of celluloseof higher plant cell walls, inhibited the cellulose formationof Acetobacter xylinum. The degree of inhibition reached 55%in the presence of 1 mM coumarin, which causes 70% inhibitionin the case of plant cellulose. (Received April 12, 1976; )     

Effects of immobilization on biomass production and acetic acid fermentation of Acetobacter aceti as a function of temperature and pH

Summary Acetobacter aceti cells were immobilized using entrapment in Ca-alginate gel and adsorption on preformed cellulose beads. The cell number within the supports showed no significant alterations on changing temperature or pH, whereas the acetic acid production was slightly increased by immobilization.     

Acetobacter xylinum生产纤维素的最适培养基成分

探索出胶醋杆菌在摇瓶培养最适培养基成分为蛋白胨浓度１．０％、酵母膏０．５％、葡萄糖浓度２．０％、柠檬酸０．１１５％、乙醇１％、Ｎａ２ＨＰＯ４０．５％,ｐＨ６．０,１６０ｒ／ｍｉｎ的条件下进行摇瓶培养,细菌纤维素最大产量为７．５５ｇ／ｌ。