赤藓糖醇高产菌株的筛选、鉴定及发酵特性的初步研究

目的：筛选出发酵性能优良的赤藓糖醇高产菌株。方法：利用含50%葡萄糖的高渗培养基筛选出耐高渗酵母,采用薄层层析和高效液相色谱分析发酵液中多元醇的组成和含量,并通过形态和生理生化试验对菌株进行初步鉴定。结果：由泰山养蜂场上采集的蜂蜜、花粉、蜂巢里筛选出13株单产赤藓糖醇的野生菌,其中7株赤藓糖醇产量高于20g/L。菌株K-23经初步鉴定为球拟酵母属,在含20%葡萄糖、1%酵母膏、0.1%尿素的发酵培养基中培养144h后赤藓糖醇的浓度达46.8g/L,转化率达23.4%。结论：所获得的菌株K-23具有一定的赤藓糖醇生产能力,且具有产物单一的优良发酵性能,为该菌株的菌种改良奠定了基础。     

球头三型孢菌TO3发酵产赤藓糖醇优化

采用响应面法优化HOG1基因敲除球头三型孢菌工程菌发酵产赤藓糖醇的工艺条件,在单因素实验的基础上,采用Box-Behnken方法进行实验方案设计,利用Design-Expert软件对结果进行二次回归分析,并进行实验验证后得到最佳发酵条件:葡萄糖200.99 g/L、酵母粉10.10 g/L、Na Cl 0.80 g/L、KH2PO40.72 g/L、Mg SO40.81 g/L,初始p H 5.0,温度30℃,转速200 r/min。在此条件下,发酵120 h后,赤藓糖醇产量可达到73.96 g/L。     

丛梗孢酵母发酵产赤藓糖醇的响应面优化

为了提高丛梗孢酵母发酵产赤藓糖醇的产量,在前期单因素实验结果的基础上,利用Plackett-Burman实验设计对影响其产赤藓糖醇的发酵条件进行评估并筛选出了影响显著的3个因素:葡萄糖、初始pH和温度.采用响应面法进行实验方案设计,利用SAS软件对其结果进行二次回归分析,确定了优化后的发酵条件为:葡萄糖260g/L、酵...     

常压室温等离子体(ARTP)诱变介导的赤藓糖醇生产菌的筛选与优化

赤藓糖醇是一种天然多元醇类甜味剂,具有热量较低、无致龋齿性、口感佳、非吸湿性以及对糖尿病患者安全等优点,已成为大多数保健食品成分的优先选择。与化学合成方法相比,赤藓糖醇的微生物发酵生产过程温和,易于控制,具有更多的生产优势。在本研究中,笔者从蜂巢、土壤和花粉等物质中筛选出了一株赤藓糖醇高产菌株,鉴定其为Moniliella sp.MU1。用常压室温等离子体(ARTP)生物育种系统对该菌株进行突变并从中挑选出4种菌株进行发酵并优化发酵条件,结果发现：不同种类的碳源、葡萄糖浓度和金属离子条件都会影响赤藓糖醇的产量,高浓度的葡萄糖有利于赤藓糖醇的生产;同时,当提供300 g/L葡萄糖和25 mmol/L Ca²⁺时,菌株YE-A27的赤藓糖醇产量达到97.6 g/L。     

新型甜味剂--赤藓糖醇产生菌的筛选

从土壤、酿造食品、花粉等样品中分离赤藓糖醇产生菌的方法,分离得到约３００株耐高渗酵母,其中有３株菌产赤藓糖醇,２６０８在葡萄糖培养液中可产赤藓糖醇３２ｍｇ／ｍｌ。     

HOG1抑制剂调节球头三型孢菌多元醇生产及机理

目的:采用HOG1抑制剂对球头三型孢菌产多元醇进行调控。方法:向培养基中加入SB239063、SB202190和SB203580三种抑制剂进行发酵实验,比较三种抑制剂对发酵的影响。结果:实验结果表明SB239063可以降低球头三型孢菌细胞内胞浆3-磷酸甘油脱氢酶(ct GPD)的酶活,提高赤藓糖还原酶(ER)的酶活。此外对HOG1和Phospho-HOG1的Western blot结果分析显示,SB239063还会抑制球头三型孢菌细胞内HOG1的脱磷酸化。最终添加10μmol/L SB239063使发酵120 h后的多元醇产物中甘油产量下降20.57%,赤藓糖醇产量提高31.16%,底物转化率提高24.73%。结论:SB239063可以降低球头三型孢菌产甘油的能力,提高赤藓糖醇的产量。     

发酵产泡性能降低的解脂耶氏酵母菌株的获得及其评价

微生物发酵过程中泡沫的产生是发酵领域遇到的共性问题。在不影响发酵性能的前提下抑制菌株的产泡,对简化操作以及降低发酵成本具有较为重要的意义。解脂耶氏酵母(Yarrowia lipolytica,之前称为Candida lipolytica)是一种常用的合成生物学底盘,也是合成赤藓糖醇等功能糖醇的生产菌株。但在发酵合成赤藓糖醇的过程中会产生大量的泡沫,需要添加消泡剂以消除泡沫。【目的】本研究旨在开发一种产泡能力显著降低的解脂耶氏酵母新菌株,以减少赤藓糖醇发酵过程中消泡剂的添加。【方法】本研究利用解脂耶氏酵母中非同源靶向重组占支配地位的原理,采用一段外源DNA随机插入基因组的手段,随机突变基因组,改变菌株的发酵产泡性能,使突变株在发酵过程中不产泡或者降低其产泡的能力。【结果】通过筛选,获得一株在发酵过程中产泡性能显著降低的工程菌株,该菌株在保留高效合成赤藓糖醇性能的同时,显著降低了泡沫的产生。【结论】所获得的菌株对工业发酵合成赤藓糖醇具有较为重要的意义,也为控制其他微生物发酵过程中泡沫的生成提供了思路。     

亚致死浓度赤藓糖醇对红火蚁工蚁觅食和弃尸行为的影响

红火蚁是国际上最具危险性外来害虫之一,已入侵华南地区并造成严重危害。为弄清潜在杀虫剂赤藓糖醇在亚致死浓度下对红火蚁的影响,本研究使用0.05 g/mL赤藓糖醇水溶液喂食室内蚁群24 h,观察了工蚁的觅食行为和弃尸行为的变化。结果显示,取食赤藓糖醇后工蚁发现食物时间约为5 min,而取食10%蔗糖水后发现食物时间缩短到2 min以内;取食赤藓糖醇后工蚁招募速度和食物搬运效率也显著低于对照。取食赤藓糖醇后工蚁发现尸体时间和弃尸速度变化显著,发现尸体时间由对照的3.45 min延长至8.13 min,开始丢弃尸体时间由20 min延长至37.5 min,丢弃完所有尸体耗时由3.08 h增大至5.37 h。亚致死浓度赤藓糖醇显著降低了红火蚁工蚁觅食和弃尸能力。     

产赤藓糖醇菌种的诱变育种

以耐高渗酵母为出发菌株,采用紫外线诱变处理,得到了一株变异株C1。其摇瓶发酵产赤藓糖醇24．9g／L左右。通过摇瓶发酵试验,研究了菌株C1的遗传稳定性。研究结果表明,经多次传代实验,菌株C1的发酵产糖醇能力没有改变,表明其是稳定的变异株。赤藓糖醇的产量由诱变前的12、594g／L提高到24.943g／L,提高了98.1％。关键词：     

赤藓糖醇对主要致龋链球菌及耐氟菌株生长和产酸的影响

目的通过赤藓糖醇对变形链球菌、远缘链球菌及其耐氟菌株混合菌生长和产酸影响的体外研究,为赤藓糖醇防龋作用的机理提供制论依据。方法采用最小抑菌浓度递增法对变形链球菌（S.mutans ATCC 25175,S.m）、远缘链球菌（S.sobrinus 6715,S.s）进行氟化钠体外诱导耐氟菌株（S.m-FR、S.s-FR）,利用液体稀释法配制赤藓糖醇TSB液8个浓度,分别加入含有变形链球菌、远缘链球菌及其耐氟菌株的细菌混悬液48 h,用比浊法观察其对混合菌生长的影响,并用pH计测定培养前后上清液的△pH值。结果吸光度A值和△pH值实验前后与对照组相比最低浓度为12%时差异均有统计学意义（P〈0.05）,且随着浓度的升高A值和△pH值均下降。结论赤藓糖醇能抑制变形链球菌、远缘链球菌及耐氟菌株混合菌生长和产酸,并且随着浓度的升高抑制作用增强。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.