葡糖醋杆菌利用黄水发酵生产细菌纤维素

为探究葡糖醋杆菌（Gluconacetobacter）利用黄水生产细菌纤维素的可行性,将黄水按不同体积比添加到传统发酵液（HS培养基）中,接种葡糖醋杆菌至发酵液中30 ℃静置培养7 d,测定发酵过程中的细菌纤维素产量、还原糖含量等理化指标。结果表明,在HS培养基中添加黄水可以显著地增加细菌纤维素的产量（P＜0.05）,黄水的最佳体积比为40%。在此条件下,细菌纤维素产量为5.93 g/L,比HS培养基（2.20 g/L）提高了169.5%;糖转化效率提高了148.9%;从第2天开始,单日细菌纤维素产量均＞0.70 g/L,第5天产量最高为1.14 g/L,单日糖转化效率均高于同时期的HS培养基;pH值维持在4.60～5.50之间。在扩大浅圆盘发酵中,40%黄水-HS培养基中细菌纤维产量为3.48 g/L,比HS培养基（1.62 g/L）提高了114.8%。     

纯菌种培养红茶菌中细菌纤维素的合成

实验采用纯菌种培养红茶菌研究细菌纤维素(bacterial cellulose,BC)的合成量及产率,所用的菌株为汉逊氏葡糖酸醋杆菌(Gluconacetobacter hansenii CGMCC1671)和啤酒酵母(Saccharomyces cerevisiae CGMCC1670),所用培养基为糖茶水培养基。研究了碳源、茶叶种类、茶叶用量、种龄、温度和装液量等6个因素对红茶菌合成BC的影响。静止培养22d后测BC的产量和产率。结果表明红茶菌合成BC的最佳碳源为葡萄糖,最佳茶叶种类为绿茶,最佳茶叶用量为4g/L,最佳种龄为60h,最佳温度为30℃,最佳装液量为250mL的三角瓶中装75mL的培养液。这些最佳参数和红茶菌的培养相一致。纯菌种培养红茶菌技术可以应用于生产高品质及高产量的BC和高品质的红茶菌饮料。     

细菌纤维素膜对木醋杆菌发酵生产广式米醋的影响

以分离自广式米醋生产车间的木醋杆菌RF4(Gluconacetobacter xylinus)为菌种进行表面发酵。研究了发酵过程中细菌纤维素膜对总酸度的影响,纤维素膜内与发酵液中乙醇脱氢酶活性差异,讨论了3种不同接种培养方式对总酸度、黏度及浑浊度的影响。结果表明,纤维素膜完整性对发酵总酸度有重要影响,纤维素膜内乙醇脱氢酶活性是发酵液中的8倍,达2.26×10-2U/g。含木醋杆菌纤维素膜接种并中途取出的接种培养方式总酸度最高,发酵12天后可达4.86 g/100 mL,且黏度及浑浊度都较低。     

不同底物的红茶菌发酵液对消化酶活性的影响

对从红茶菌中分离出的菌株(Gluconacetobacter sp.A4)进行研究,分别以红茶、荷叶、荷叶与山楂、荷叶与决明子为底物进行发酵,以传统红茶菌混合发酵为对照,通过对其产物D-葡萄糖二酸-1,4-内酯含量等指标的测定来评价发酵效果,同时研究了不同底物红茶菌发酵液对消化酶活性的影响。结果表明:在30℃、发酵8 d的情况下,以荷叶山楂A4发酵液发酵最为充分,其次为红茶A4发酵液。红茶A4发酵液的D-葡萄糖二酸-1,4-内酯(DSL)生成量最高。各发酵液对脂肪酶的抑制作用以红茶A4发酵液抑制作用最强,荷叶A4发酵液次之;对α-淀粉酶的抑制作用,以荷叶决明子A4发酵液的抑制率最高,红茶A4发酵液次之。不同红茶菌发酵液对消化酶的活性具有抑制作用,为其减肥机理的研究以及功能饮料的开发奠定基础。     

以柑橘果渣为主要原料生产细菌纤维素的研究

目的:拓宽细菌纤维素(BC)生产的原料来源,明确柑橘渣水对汉逊氏葡糖酸醋杆菌CIs51发酵产BC的影响。方法:以筛选自酸败柑橘果实的汉逊氏葡糖酸醋杆菌CIs51为发酵菌株,以柑橘果渣为主要原料,用扫描电镜观察其微结构,研究果渣预处理工艺、营养源、生长因子等对BC产量及微结构的影响。结果:柑橘果渣与水以1:8(W/V)的比例混合打浆,以150U/mL的果胶酶复配100U/mL纤维素酶,45℃水解2h;过滤后进行酵母前发酵工艺;选择蔗糖为碳源,添加量30g/L,(NH4)2SO4为氮源,添加量3g/L,酵母粉和柠檬酸的添加量分别为5g/L和1g/L。在此优化培养基中CIs51的产量达7.23g/L,明显高于其在HS培养基、柑橘渣水改良HS(CMHS)培养基中的产量(依次为4.02g/L及6.57g/L)。结论:柑橘渣水能够显著提高CIs51的BC产量,所得BC膜呈半透明质地,柔韧性好,具有替代椰子水进行工业化生产的前景。     

Effects of Lactobacillus casei and Alterations in Fermentation Conditions on Biosynthesis of Glucuronic Acid by a Dekkera bruxellensis-Gluconacetobacter intermedius Kombucha Symbiosis Model System

A combination of Dekkera bruxellensis and Gluconacetobacter intermedius is used as a model system to emulate the microbial symbiosis between yeast and acetic acid bacteria in kombucha, and to study glucuronic acid (GlcUA) production in this system. In the current study, the addition of Lactobacillus casei and variations in the fermentative conditions, such as sucrose concentration, temperature and duration, were examined to evaluate their effects on GlcUA formation. Results showed that L. casei acts as a supporter species and stimulated GlcUA production in the kombucha symbiosis model to levels 54.1% greater than in the control. Varying the fermentation parameters also affected GlcUA production. Therefore, the range of each factor value was screened and restricted to achieve a higher amount of target acid, as it was 5–15%, 25–31°C, 3–7 days, and 5–7 units in sucrose concentration, temperature, fermentation time and initial pH value, respectively. These findings are an important step for optimizing GlcUA production by this model system.     

HPLC法检测红茶菌中葡萄糖代谢曲线及培养基的优化研究

本实验室从红茶菌中分离出可以产生高含量的功能因子-葡萄糖二酸1,4内酯的菌株--葡糖酸醋酸杆菌(Gluconacetobactersp.A22)。本文利用高效液相色谱法检测葡萄糖,通过对葡萄糖代谢曲线的测定,确定了发酵时间为126h(30℃,160r/min振荡培养)。发酵结束时,残糖量与初始葡萄糖浓度呈线性关系:y=40.254x-35.862(R2=0.9999)。对A22培养基的优化进行了研究,结果表明,葡萄糖浓度5%(w/v)、茶叶浓度0.5%(w/v)时最适合A22发酵生产功能性饮料。     

The Oxidative Fermentation of Ethanol in Gluconacetobacter diazotrophicus Is a Two-Step Pathway Catalyzed by a Single Enzyme: Alcohol-Aldehyde Dehydrogenase (ADHa)

Gluconacetobacter diazotrophicus is a N₂-fixing bacterium endophyte from sugar cane. The oxidation of ethanol to acetic acid of this organism takes place in the periplasmic space, and this reaction is catalyzed by two membrane-bound enzymes complexes: the alcohol dehydrogenase (ADH) and the aldehyde dehydrogenase (ALDH). We present strong evidence showing that the well-known membrane-bound Alcohol dehydrogenase (ADHa) of Ga. diazotrophicus is indeed a double function enzyme, which is able to use primary alcohols (C2–C6) and its respective aldehydes as alternate substrates. Moreover, the enzyme utilizes ethanol as a substrate in a reaction mechanism where this is subjected to a two-step oxidation process to produce acetic acid without releasing the acetaldehyde intermediary to the media. Moreover, we propose a mechanism that, under physiological conditions, might permit a massive conversion of ethanol to acetic acid, as usually occurs in the acetic acid bacteria, but without the transient accumulation of the highly toxic acetaldehyde.