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。     

The terminal protein of the linear DNA plasmid pGKL2 shares an N-terminal domain of the plasmid-encoded DNA polymerase

The 36K protein attached at the 5′ end of the linear DNA plasmid pGKL2 from the yeast Kluyveromyces lactis was first purified and characterized. The terminal protein was purified from cells (1 kg wet weight) by ammonium sulphate precipitation and two rounds of centrifugation to equilibrium in CsCl gradients. The pGKL2 was present only in the post-microsomal supernatant. Approximately 10 mg of the purified pGKL2 was recovered and digested with DNase I. The terminal protein (final ca. 0·8 mg) was homogeneous by electrophoresis and we determined the N-terminal amino acid sequence up to ten residues, showing that it existed in the cryptic N-terminal domain of pGKL2-ORF2 (DNA polymerase) sequence.     

食品生物防腐剂产生菌门间远缘细胞融合研究

乳酸链球菌素(Nisin)和ε-聚赖氨酸是由微生物产生的安全的天然防腐剂,分别由乳酸链球菌和白色链霉菌产生。这两种生物防腐剂的抑菌效果不同,存在互补性。为了获得具有更广谱抑菌作用的优良性状的菌株,探索远缘细胞融合的可行性,以自主分离的ε-聚赖氨酸产生菌白色链霉菌YB12和Nisin产生菌NZ9700作为亲本菌株,通过原生质体融合技术将两个菌株进行门间的细胞融合;通过抗药性筛选、形态学观察、SRAP-PCR分子鉴定,成功获得两株稳定遗传的融合菌株。同时对融合菌株的抑菌活性进行了检测。     

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.     

乳酸乳球菌NICE系统食品级载体pRNB48的构建

乳酸菌食品级NICE系统是目前较理想的可控制蛋白质生产的食品级诱导表达系统。实验构建的食品级表达载体pRNB48含α-aga食品级选择标记、θ型复制子和nisA启动子,与宿主菌L.lactisNZ9000共同构成乳酸乳球菌食品级NICE系统,可用于目的基因在乳酸乳球菌中高效诱导表达。     

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。