Enhanced production of extracellular inulinase by the yeast Kluyveromyces marxianus in xylose catabolic state

1. Download high-res image (194KB)
2. Download full-size image

     

Efficient production of L-lactic acid from xylose by a recombinant Candida utilis strain

Efficient L-lactic acid production from xylose was achieved using a pyruvate decarboxylase-deficient Candida utilis strain expressing an L-lactate dehydrogenase, an NADH-preferring mutated xylose reductase (XR), a xylitol dehydrogenase and a xylulokinase. The recombinant strain showed 53% increased L-lactic acid production compared with the reference strain expressing native XR (NADPH-preferring).     

Favorable effect of very low initial KLa value on xylitol production from xylose by a self-isolated strain of Pichia guilliermondii

Xylitol production from xylose by a self-isolated furfural and 5-hydroxymethyl furfural assimilating Pichia guilliermondii was studied under oxygen limitation. An extremely low initial volumetric oxygen transfer coefficient (0.075 h^− 1) was found most favorable to the xylitol production with yield of 0.61 g g^− 1. Related enzymes activities were also investigated and discussed.     

Sucrose hydrolysis by gelatin-immobilized inulinase from Kluyveromyces marxianus var. bulgaricus

The crude cell-free medium from a culture of Kluyveromyces marxianus var. bulgaricus was immobilized in a gelatin-water support, with an immobilization yield of 82.60% for inulinase activity. The optimum pH for both free and immobilized inulinase was the same (3.5) and the optimum temperatures were 55 °C for the free and 60 °C for the immobilized enzyme. The Arrhenius plots were linear and activation energies were 56.20 (free enzyme) and 20.27 kJ/mol K (immobilized enzyme). The kinetic parameters were calculated by Lineweaver–Burk plots and the V_max and K_m were 37.60 IU/mg protein and 61.83 mM for the free inulinase and 31.45 IU/mg protein and 149.28 mM for the immobilized enzyme, respectively. The operational stability of the immobilized inulinase was studied in a continuous fixed-bed column reactor for 33 days, at the end of which the sucrose conversion was 58.12%.     

Evaluation of thermally-dried Kluyveromyces marxianus as baker’s yeast

The aim of the present study was to evaluate the use of thermally-dried Kluyveromyces marxianus as baker’s yeast. Bread samples produced by thermally-dried K. marxianus were compared with samples produced by wet K. marxianus culture and by commercial baker’s yeast. The type of the culture had no effect on loaf weight, loaf volume, specific loaf volume, density or moisture loss, in contrast to pH, total titrable acidity (TTA), and moisture content. The use of thermally-dried K. marxianus resulted in lower pH values and higher TTA, while the bread samples showed higher resistance to spoilage as counts of moulds and yeasts were significantly lower during preservation. The SPME GC/MS analysis of volatiles and the preliminary sensory evaluation showed no significant differences in the profile of aroma-related compounds or overall quality of the tested samples.     

Influencing cocoa flavour using Pichia kluyveri and Kluyveromyces marxianus in a defined mixed starter culture for cocoa fermentation

The potential impact of aromatic and pectinolytic yeasts on cocoa flavour was investigated using two defined mixed starter cultures encompassing strains of Pichia kluyveri and Kluyveromyces marxianus for inoculating cocoa beans in small scale tray fermentations. Samples for microbial and metabolite analysis were collected at 12–24 hour intervals during 120 h of fermentation. Yeast isolates were grouped by (GTG)₅-based rep-PCR fingerprinting and identified by sequencing of the D1/D2 region of the 26S rRNA gene and the actin gene. Pulsed Field Gel Electrophoresis (PFGE) was conducted on isolates belonging to the species P. kluyveri and K. marxianus to verify strain level identity with the inoculated strains. Furthermore, Denaturing Gradient Gel Electrophoresis (DGGE) was performed to follow yeast and bacterial dynamics over time including the presence of the bacterial inoculum consisting of Lactobacillus fermentum and Acetobacter pasteurianus. Yeast cell counts peaked after 12 h of fermentation with the predominant species being identified as Hanseniaspora opuntiae and Hanseniaspora thailandica. P. kluyveri and K. marxianus were found to compose 9.3% and 13.5% of the yeast population, respectively, after 12 h of fermentation whilst PFGE showed that ~ 88% of all P. kluyveri isolates and 100% of all K. marxianus isolates were identical to the inoculated strains. Despite never being the dominant yeast species at any stage of fermentation, the un-conched chocolates produced from the two inoculated fermentations were judged by sensory analysis to differ in flavour profile compared to the spontaneously fermented control. This could indicate that yeasts have a greater impact on the sensory qualities of cocoa than previously assumed.     

混合糖发酵产油脂酵母菌的筛选

对6株产油脂酵母利用葡萄糖、木糖及不同比例混合糖(葡萄糖与木糖)发酵的菌体生长和产油特性进行研究,筛选到一株能够转化混合糖的高产油脂酵母JM-D.该菌株在混合糖比例为3∶1的条件下发酵120h,菌体油脂含量可达23.38％,油脂产量可达2.7g/L;菌体油脂中饱和脂肪酸含量占脂肪酸总量的49.2％;混合糖比葡萄糖更容易被转化生成饱和脂肪酸含量较多的油脂.     

以L-苏氨酸为发酵底物的2,5-二甲基吡嗪高产菌株构建

构建一种以L-苏氨酸为发酵底物的高值化学品2,5-二甲基吡嗪(2,5-dimethylpyrazine, 2,5-DMP)生产菌株,为解决L-苏氨酸产能过剩,实现2,5-DMP生物法生产提供可靠思路。通过利用Bacillus subtilis 168(B.subtilis 168)外源表达不同微生物种属来源的L-苏氨酸脱氢酶(L-threonine dehydrogenase, TDH),并比较其利用L-苏氨酸为底物合成2,5-DMP的产量,挑选出2,5-DMP高产菌种,在此基础上进一步外源表达NADH氧化酶(NADH oxidase, NOX),以促进辅因子再生。实验构建了1株高产2,5-DMP的基因工程菌株B.subtilis 168/pMA0911-tdh(E.c)-nox。该菌株以5.83 g/L的L-苏氨酸为底物,发酵24 h后2,5-DMP的产量高达616.04 mg/L,与对照菌株B.subtilis 168/pMA0911相比,产量提高了22.5倍。在TDH过表达的基础上,NOX的参与有利于2,5-DMP产量的提高。该研究首次实现了2,5-DMP高效的生物转化,一方面缓...     

Application of Kluyveromyces marxianus, Lactobacillus delbrueckii ssp. bulgaricus and L. helveticus for sourdough bread making

The application of Kluyveromyces marxianus (IFO 288), Lactobacillus delbrueckii ssp. bulgaricus (ATCC 11842) and Lactobacillus helveticus (ATCC 15009) as starter cultures for sourdough bread making was examined. Production of lactic and acetic acids, bread rising, volatile composition, shelf-life and organoleptic quality of the sourdough breads were evaluated. The amount of starter culture added to the flour, the dough fermentation temperature and the amount of sourdough used were examined in order to optimise the bread making process. The use of mixed cultures led to higher total titratable acidities and lactic acid concentrations compared to traditionally made breads. Highest acidity (3.41 g lactic acid/kg of bread) and highest resistance to mould spoilage were observed when bread was made using 50% sourdough containing 1% K. marxianus and 4% L. delbrueckii ssp. bulgaricus. The use of these cultures also improved the aroma of sourdough breads, as shown by sensory evaluations and as revealed by GC–MS analysis.     

Evolution of aroma volatiles during storage of sourdough breads made by mixed cultures of Kluyveromyces marxianus and Lactobacillus delbrueckii ssp. bulgaricus or Lactobacillus helveticus

Two mixed starter cultures were used for sourdough bread making to evaluate their ability to improve quality and increase bread shelf-life: Lactobacillus delbrueckii ssp. bulgaricus or Lactobacillus helveticus mixed with the lactose fermenting yeast Kluyveromyces marxianus as alternative baker’s yeast. Control sourdough breads (K. marxianus) without the addition of bacteria, were also prepared. The changes on the headspace aroma volatiles during storage were assessed using solid-phase microextraction (SPME) GC–MS analysis. The effect of these changes on bread flavour was evaluated by consumer preference evaluations and the results were co-evaluated with those from the GC–MS analysis. The obtained results showed differences in the volatile composition of the different types of breads examined, as well as dramatic decreases of the number and the amount of volatiles after five days of storage. The sourdough breads made with K. marxianus and L. bulgaricus, had a more complex aroma profile, longer shelf-life and achieved the highest scores in the sensory tests.     

直接发酵木糖为酒精的新酵母基因工程菌的选育

裂殖酵母,Schizosacchatomyces porebe不能发酵木糖,也不能用它来增殖细胞。但是,这种酵母能发酵木酮糖为酒精,产率和转化率都很高。这表明,它缺少异构酶基因,如果将其它微生物的这种基因克隆到酵母中去,它就能直接发酵木糖。本文介绍了Schizosaceh.porebe的五碳糖磷酸代谢途径,这种代谢途径使得它成为用于这一目的的理想菌株。含有异构酶基因的DNA片段来自Clark和Carbon的大肠杆菌基因库,它的大小为2.4kb,PstⅠ-抗性。将这个基因克隆到上述酵母中去,得到的酵母基因工程菌株不仅能利用木糖作为唯一碳源和能源,还能发酵木糖为酒精,它具有在工业上应用的潜力     

高浓度蔗糖酒精发酵的初步研究

高产量酒度和快速发酵一直是酒精发酵产业所追求的,而酵母直接利用高浓度蔗糖发酵酒精未见有文献报道.文中所用酿酒酵母能直接发酵30%(w/v)的蔗糖溶液,发酵周期只有2d～3d,其最终产生的酒精可以达到17.08%vol,对发展蔗糖酒精是个很好的契机.     

酵母发酵法去除木糖母液中葡萄糖的研究

以木糖母液为原料,采用酵母发酵法去除母液中的葡萄糖。通过单因素分析,对影响去除效果的主要因素进行了研究。利用正交试验筛选出最佳工艺参数。结果表明:母液浓度为12%,调pH值4.5,酵母添加量2%,30℃下发酵6h,葡萄糖的去除率可达96%以上。     

A comparative study of ethanol production by Issatchenkia orientalis strains under stress conditions

The ability of 13 strains of multi-stress-tolerant Issatchenkia orientalis yeast to produce ethanol was examined under different stress conditions, including conditions of elevated H₂SO₄ and Na₂SO₄ concentrations and increased heat. The MF-121 strain produced a significant amount of ethanol after the incubation in acidic media containing high concentrations of salt, e.g., 50 g/l Na₂SO₄ at pH 2.0, or at high temperatures, e.g., 43°C, when compared with other strains.     

In vitro fermentation of xylo-oligosaccharides from corn cobs autohydrolysis by Bifidobacterium and Lactobacillus strains

The selective release of soluble oligomeric structures from corn cobs (CC) by autohydrolysis was optimized using the severity factor (log R₀), and a maximum xylo-oligosaccharides (XOS) production was obtained at log R₀=3.75. Gel filtration chromatography was employed to separate oligosaccharides (OS) from the crude hydrolysate and two fractions with average degrees of polymerization (DP) of 3-4 and 5-6 were compared with commercial XOS in their ability to promote the growth of Bifidobacterium adolescentis, B. longum, Lactobacillus brevis and L. fermentum as carbon and energy sources. Among the tested strains, B. adolescentis and L. brevis displayed the highest growth and XOS consumption, in contrast to B. longum and L. fermentum whose overall growth on XOS was low. XOS mixtures from CC autohydrolysis, mainly constituted by xylotriose and xylotetraose, were fermented by B. adolescentis as well as commercial XOS containing essentially xylobiose, whereas L. brevis preferred XOS with an average DP of 2. However, an increase in XOS chain length to DP 5-6 clearly reduced the extension to which B. adolescentis utilized these OS. This study showed that XOS of low molecular weight from CC autohydrolysis exhibit a potential bifidogenic capability similar to commercial XOS.