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。     

Production of human growth hormone by Lactococcus lactis

A synthetic gene coding for human growth hormone was expressed in Lactococcus lactis. The presence of the recombinant protein was assayed and quantified using ELISA tests. Human growth hormone was detected at high concentrations and displayed a biological activity similar to the one shown by commercial human growth hormone.     

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。     

Environmental factors for phage-induced fermentation problems: replication and adsorption of the Lactococcus lactis phage P008 as influenced by temperature and pH

The lactococcal phage P008 was investigated for its growth characteristics under certain environmental conditions. Phage growth was characterized by the latency period, the average burst size (determined in one-step growth experiments) and by the percentage of adsorption to the host cells after 1, 5, 10 and 15 min (determined in modified one-step growth experiments). The incubation conditions studied were temperatures from 20 to 40 degrees C and pH values of 4.8 and 6.4. The growth medium was ultrafiltration permeate of skim milk, which forms the water phase of milk. Both, the temperature and the pH influenced the growth parameters: Increased temperature as well as low pH led to a faster latency along with a higher average burst size. The percentage of adsorption was at maximum when the standard conditions of 30 degrees C and pH 6.4 were applied. Below pH 5, adsorption was reduced to below 10%. A decrease of temperature slightly reduced phage multiplication. Data were incorporated into a model to simulate phage multiplication under certain conditions, taking the percentage adsorption into account. The cell destruction of the host culture was calculated accordingly. The simulation (extrapolation) was validated by experimental growth curves of phages and phage-infected host cultures.     

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。     

一株瑞士乳杆菌增殖培养基的优化分析

通过单因素实验、正交实验研究了不同碳源、氮源、碳氮比例、缓冲盐浓度、微量元素及其他生长因子对瑞士乳杆菌(Lactobacillushelveticus H9)增殖培养的影响。并采用响应面法对优选的碳源、氮源和缓冲盐类的组成质量浓度进行优化,得到的增殖培养基:葡萄糖20 g/L,大豆蛋白胨10 g/L,酵母粉5 g/L,NaAC为7 g/L,K2HPO4为3 g/L,柠檬酸钠1 g/L,MgSO4.7H2O为200 mg/L、MnSO4.5H2O为81 mg/L、L-半胱氨酸0.25 g/L、吐温80为1 g/L。Lactobacillus helveticus H9在此增殖培养基中经37℃,10 h培养活菌数可达到1.34×1010mL-1,比在普通MRS中提高近35倍。     

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。