温度对透明质酸分批发酵的影响及动力学模型

文章研究了30℃－40℃范围内透明质酸（HA）分批发酵过程,发现最适的细胞生长温度是34℃,而40℃时HA的产量最高为0.810g.L^-1。以现有的底物抑制和产物抑制动力学模型为基础,建立HA发酵过程中菌体生长,HA合成,乳酸合成及葡萄糖消耗的动力学模型,并计算出不同温度下相关的动力学参数,在此基础上,进一步研究了温度同细胞生长动力学参数之间的内在联系,结果表明H inshelwood模型能够很好地反应在HA分批发酵过程中细胞浓度的变化同温度以及底物浓度之间的一般关系式。     

Lactobacillus delbrueckii subsp. bulgaricus和Streptococcus thermophilus接种比例对Bifidobacterium lactis益生菌发酵乳品质的影响

将Lactobacillus delbrueckii subsp.bulgaricus ND02(LB-ND02)和Streptococcus thermophilus ND03(ST-ND03)按1∶1、1∶10、1∶100、1∶1000接种于脱脂乳中,同时接入益生菌Bifidobacterium lactis V9(B.lactis V9,接种量为2.0×107g-1),于42℃进行发酵。通过对发酵及贮藏过程中发酵乳指标的测定,评价LB-ND02和ST-ND03的接种比例对发酵乳品质的影响。结果表明,随着LB-ND02接种比例减小,凝乳时间显著延长,B.lactis V9活菌数显著提高。4℃贮藏28 d后,随LB-ND02接种比例减小,B.lactis V9存活率差异显著,后酸化也显著减弱。研究发现,LB-ND02和ST-ND03的接种比例,显著影响发酵乳的发酵时间、B.lactis V9活菌数、后酸化及黏度。     

A群链球菌四价重组蛋白的表达、纯化及其免疫原性

目的原核表达并纯化A群链球菌M1、3、6、18型四价重组蛋白,并检测其免疫原性。方法以克隆质粒pUC18-Strep4为模板,PCR法扩增四价重组蛋白基因,克隆至表达载体pQE30中,构建重组表达质粒,转化E.coilM15,筛选阳性克隆,IPTG诱导表达。采用Ni2+-NTA凝胶亲和层析纯化重组蛋白。以该蛋白免疫家兔,ELISA法检测血清抗体滴度;间接免疫荧光法(IFA)检测A群链球菌型特异性抗体;体外杀菌试验检测杀菌抗体活性。结果四价重组蛋白表达质粒pQE30-Strep4经双酶切和测序,证明构建正确。重组蛋白相对分子质量与预期相符,表达量约占菌体总蛋白的30%,为可溶性表达。纯化后蛋白纯度可达90%以上,并可诱导家兔产生高滴度的M1、3、6型特异性杀菌抗体。结论已成功表达了A群链球菌四价重组蛋白,纯化后的蛋白可诱导家兔产生针对A群链球菌M1、3、6三个血清型菌株的特异性杀菌抗体。     

Nucleotide sequence of the cytochrome oxidase subunit 2 and val-tRNA genes and surrounding sequences from Kluyveromyces lactis K8 mitochondrial DNA

The nucleotide sequence of the cytochrome oxidase subunit 2 (cox2) and val-tRNA genes and surrounding regions from Kluyveromyces lactis mitochondrial DNA is reported. Analysis of the coding region shows that the codons CUN (Thr), CGN (Arg) and AUA (Met) are absent in this gene. A single sequence, ATATAAGTAA, identical to the baker's yeast mtRNA polymerase recognition site, was detected upstream of val-tRNA. This sequence is absent from regions between val-tRNA-cox2 and cox2-cox1. In addition a sequence AATAATATTCTT, identical to the mRNA processing site in other yeast mitochondrial genomes is present 32-43 bp downstream to the TAA stop codon for the cox2 gene. Another short conserved sequence of 5 bp, TCTAA, is present upstream of the coding regions of cox2 genes in several yeasts, including K. lactis, but is not present upstream of other genes. Comparison of cox2 sequences from other organisms indicates that the mitochondrial DNA of K. lactis is closely related to that of Saccharomyces cerevisiae.     

cA-GFP融合蛋白在大肠杆菌中的高效功能性表达

为了实现乳酸乳球菌锚定蛋白cA在大肠杆菌中可溶表达以及直观检测其生物学活性,以乳酸乳球菌MG1363为模板,扩增N-乙酰葡萄糖胺糖苷酶AcmA基因,将其C末端序列与GFP基因融合,连接至大肠杆菌表达载体pET28a,构建重组质粒pET28a-cA-GFP,转化至大肠杆菌表达菌株BL21(DE3),通过低温诱导表达重组蛋白cA-GFP。工程菌超声破碎后的上清液与经热酸处理的乳酸乳球菌常温孵育,经SDS-PAGE和荧光显微镜检测。结果表明,工程菌可以表达分子量约53ku的目的融合蛋白质,与预期大小相符。cA-GFP通过锚定蛋白cA的引导回向锚定,成功将GFP展示在乳酸乳球菌表面,目的蛋白cA-GFP在乳酸乳球菌表面展示量为121mg/g干重菌体。GFP锚定至乳酸乳球菌后于4℃保存,连续6d测定其荧光强度,荧光强度仍可达82.2%,证明其稳定性较好。成功获得了具有生物学功能的cA-GFP可溶性蛋白,为进一步展示功能性外源蛋白奠定了坚实的基础。     

腐乳源乳酸乳球菌17M1高密度培养条件研究

为了获得乳酸乳球菌(Lactococcus lactis)17M1的高密度培养方法,以菌体密度和活菌数为评价指标,以豆腐乳清为基础培养基,采用响应面法对该菌株的培养基组成进行优化,并研究其培养条件。结果表明,菌株17M1的最佳培养基组成为：在豆腐乳清中加入2.00%大豆蛋白胨、胰酪蛋白胨1.70%、柠檬酸铵2.30%。在此优化培养基中初始pH值6.15,于37℃培养15 h,乳酸乳球菌17M1的活菌数达到了1.17×10¹⁰CFU/mL,高于M17肉汤培养基活菌数(1.04×10⁹CFU/mL)。该研究结果为乳酸乳球菌17M1的工业化应用提供了技术支持。     

Probiotic potential and sensory properties of coconut flan supplemented with Lactobacillus paracasei and Bifidobacterium lactis

The effect of probiotic cultures on sensory performance of coconut flan during storage at 5 °C and the viability of these micro organisms for up to 28 days were investigated. Sensory analyses of the product were performed after 7, 14 and 21 days of storage. Coconut flans were produced with no addition of cultures (T1, control), or supplemented with Bifidobacterium lactis (T2), Lactobacillus paracasei (T3) and B. lactis  +  L. paracasei (T4). Populations of L. paracasei and B. lactis as single or in co-culture remained above 7 log CFU g⁻¹ during the entire storage period. Viability of L. paracasei was higher for T3. All products were well accepted and no significant differences ( P  > 0.05) were detected between the coconut flans studied. The addition of L. paracasei and B. lactis to coconut flan resulted in its having great potential as a functional food, which has high sensory acceptability.     

LYS2 gene and its mutation in Kluyveromyces lactis

The KlLYS2 gene, encoding the alpha-aminoadipate reductase of Kluyveromyces lactis, was isolated by complementation of a lysA1 mutant. The deduced amino acid sequence shared an identity of 73% with the LYS2 product of Saccharomyces cerevisiae. Despite the high sequence homology of the alpha-aminoadipate reductase genes, the two yeast species differently responded to the presence of alpha-aminoadipate in the medium. Wild-type S. cerevisiae is known to be sensitive to alpha-aminoadipate, but becomes resistant when mutated to lys2. In contrast, K. lactis strains were found to be naturally resistant to alpha-aminoadipate. Therefore, the positive selection procedure for the isolation of lys2 mutants on alpha-aminoadipate, as practised in S. cerevisiae, cannot be applied to K. lactis. A possible reason of this difference may be that the catalytic rate of the alpha-aminoadipate reductase differs in the two yeasts. The EMBL/Genbank Accession No. for the KlLYS2 gene is AJ504405.     

低温长时间发酵酸乳加工关键技术及品质研究

选用乳酸链球菌和保加利亚乳杆菌制作低温长时间发酵酸乳,利用DPS软件设计正交实验优化酸乳生产工艺,并对发酵结束和经4℃、后熟24h后以及4℃、冷藏3d后酸乳的酸度、pH值、纯蛋白、游离氨基酸、脂肪、双乙酰、乙醛、乳酸菌活菌数量等质量指标和风味成分进行分析检测。结果表明,最佳工艺为乳酸链球菌和保加利亚乳杆菌二4∶1,接种量3%,添加蔗糖8%,34℃培养10h左右。低温长时间发酵酸乳的酸度为69·35°T,乙醛含量为14·46mg/kg,双乙酰含量为4·72mg/kg,乳酸菌活菌数为7·15×109cfu/mL。经感官评定,酸乳的凝乳组织状态均匀细腻,香气浓郁纯正,酸甜适口,品质优良。