pH对Themoanaerobacterium calidifontis Rx1乙醇发酵代谢的影响

Themoanaerobacterium calidifontis Rx1是一株能降解半纤维素发酵乙醇的高温厌氧菌。本研究以Rx1和其乙酸激酶基因(ack)敲除的突变菌株Rx1△ack作为研究对象,研究了不同pH值条件对两种菌株生长与代谢的影响,并通过测定碳中心代谢关键酶的表达量与酶活性分析了pH的代谢调控机制。结果表明,与自然pH相比,控制pH能够改变两种菌株的代谢产物分布;当控制pH值为7.0时,Rx1和Rx1△ack的乳酸产量分别下降了85.7%和89.9%,而乙醇产量分别提高了60%和69%;Rx1和Rx1△ack乙酸产量的变化趋势不同。与自然pH相比,控制p H时,Rx1和Rx1△ack的丙酮酸激酶(PK)基因表达量和酶活性都有所提高;野生型菌株的乙酸激酶(ACK)活性下降11%,而突变菌株基因表达量显著提高,酶活性只有野生型的25%,且与自然pH条件相比,有提高的的趋势;野生型菌株乙醇代谢途径中乙醇脱氢酶(ADH)活性显著下调;研究发现pH对Rx1和Rx1△ack乳酸代谢途径中乳酸脱氢酶(LDH)影响十分显著,在弱碱或中性条件下,其活性还不足酸性条件下的一半。Rx1和Rx1△ack在控制pH条件下乳酸产量下降,同时伴随乳酸生成途径的还原力(即NAD(P)H)积累,而乙醇生成途径可将积累的NAD(P)H氧化成NAD(P)+,这可能是控制pH条件下乙醇产量提高的原因。     

Kluyveromyces marxianus发酵己糖和戊糖生产乙醇

对利用底物广泛的乙醇发酵菌株马克斯克鲁维(Kluyveromyces marxianus)DL1菌株与工业用乙醇发酵菌株酿酒酵母(Saccharomyces cerevisiae)6525利用己糖(葡萄糖、甘露糖、半乳糖)和戊糖(木糖、阿拉伯糖)的情况进行对比研究。结果发现:以己糖为底物时,K.marxianus DL1均表现出细胞生长快、乙醇得率高的特点;在不通气、糖20 g/L条件下,K.marxianus DL1的最大乙醇质量浓度均比S.cerevisiae 6525高出10%左右,细胞量及乙醇生产强度分别是S.cerevisiae 6525的近2和1.7倍。当以戊糖为底物时,K.marxianus DL1可以利用木糖和阿拉伯糖;在不通气、糖20g/L条件下,K.marxianus DL1利用木糖产木糖醇和乙醇,乙醇终质量浓度可达7.68 g/L,木糖醇质量浓度为9.12 g/L;以阿拉伯糖为发酵底物时,阿拉伯糖醇的产量可达6 g/L左右;而S.cerevisiae 6525不能利用戊糖。马克斯克鲁维酵母比酿酒酵母更适合纤维乙醇生产。     

氦氖激光和亚硝基胍复合诱变选育高产乙醇的休哈塔假丝酵母

木糖的乙醇发酵一直被视为木质纤维原料生物转化产生乙醇的关键因素,休哈塔假丝酵母(Candidashehatae)是木糖发酵性能较好的天然酵母之一。对Candida shehatae HDYXHT-01进行了氦氖激光诱变和NTG诱变,力求选育出发酵木糖产乙醇能力强的菌株。氦氖激光诱变得到的突变株HN-3乙醇产量为17.03g/L,乙醇得率达到0.3393g/g,相比原始菌株提高20.36%。再对HN-3进行NTG诱变,得到的突变株NTG-2乙醇产量为24.20g/L,相比HN-3提高42.10%。进而对NTG-2菌株进行了摇瓶48h连续发酵试验,测得其乙醇产量、木糖利用率、乙醇得率和乙醇产率分别达到24.16g/L,69.26%,0.4360g/g和0.7075g/(L·h)。     

玉米秸秆水解液发酵联产氢气与2,3-丁二醇的初步研究

选用实验室自行筛选的Klebsiella pneumoniae ECU-15,进行了玉米秸秆水解液发酵联产氢气和2,3-丁二醇的初步研究。结果表明:以葡萄糖为碳源时,两目标产物随培养条件的改变呈现相同的变化趋势,且最佳发酵温度为37℃,最佳pH为6.0,最佳初始糖浓度为30 g/L;不同比例葡萄糖/木糖为混合碳源时,均能实现氢气和2,3-丁二醇的联产过程,但随着木糖含量的增加,细胞产量、氢气产量和2,3-丁二醇的产量都有所下降,并且木糖的存在会降低葡萄糖的消耗速率;实验最后以玉米秸秆水解液和同比例模拟合成培养基为底物,初步探明了该菌株利用水解液发酵联产氢气和2,3-丁二醇的可行性,最终氢气产量为0.65 v/v,产氢得率为0.43 mol/mol sugar;2,3-丁二醇产量为5.05 g/L,得率为0.82 mol/mol sugar。     

亚硫酸盐甘蔗渣浆酶解液发酵制备乙醇

以亚硫酸盐甘蔗渣浆酶解液作为原料,利用C. shehatae发酵制取燃料乙醇。结果表明：还原糖最适初始质量浓度为葡萄糖140 g/L、木糖60 g/L、酶解液总糖80 g/L。利用初始葡萄糖55.06 g/L、木糖11.18 g/L、纤维二糖4.51 g/L的亚硫酸盐甘蔗渣浆酶解液发酵,经18 h获得乙醇22.98 g/L。乙醇得率为67.23%,葡萄糖利用率为99.27%,木糖利用率为32.96%,C. shehatae适合作为蔗渣为原料的乙醇发酵菌株。     

一株厌氧嗜热杆菌生长及发酵特性的初步研究

论文对筛选并鉴定为Thermoanaerobacterium saccharolyticum菌株的生长、底物利用情况、产物生成以及酒精耐受性进行了研究。结果表明,该菌在以甘露糖、葡萄糖和木糖为碳源时生长较好,同时能够较好的利用木聚糖和木薯淀粉;最适底物浓度为15g/L;不同的葡萄糖:木糖比例对其生长无显著影响;能耐受的培养基最高初始酒精浓度为3%(V/V)。在5g/L的木聚糖、木糖和木薯淀粉培养基中发酵60h后,产物主要有乙醇、乳酸和乙酸,乙醇产量分别为0.824、0.867和0.916g/L。     

玉米芯稀酸水解及L-乳酸发酵研究

对玉米芯稀硫酸水解条件及糖化液发酵L-乳酸进行了初步研究。结果表明,玉米芯木聚糖最适水解条件为2%H2SO_4、120℃、30 min、固液比1:10,糖化液还原糖含量可达40.8 g/L,主要成分为木塘。细菌A-19可以利用水解液中的葡萄糖和木糖产酸,最适发酵条件为45℃、pH 6.5,从45℃～51℃、pH 5.5～pH 6.5产量均较高。用未浓缩的水解液发酵24 h,L-乳酸产量为30.6g/L,残糖为1.6 g/L,糖酸转化率为82.6%;用浓缩1倍的水解液发酵48 h,L-乳酸产量为41.4 g/L,残糖4.1g/L,糖酸转化率为68.2%,在发酵48 h后继续补料发酵至72 h(补料液为浓缩3倍的水解液),L-乳酸产量为50.9 g/L,残糖6.3 g/L,糖酸转化率为71.8%。该研究为利用木质纤维素生产L-乳酸奠定了一定基础。     

休哈塔假丝酵母乙醇发酵性能的研究

木糖的高效发酵是制约纤维素燃料乙醇生产的技术瓶颈之一,高性能发酵菌种的开发是本领域研究的重点。以木糖发酵的典型菌株休哈塔假丝酵母为材料,研究氮源配比、葡萄糖和木糖初始浓度、葡萄糖添加及典型抑制物等因素对其木糖利用和乙醇发酵性能的影响规律。结果表明,硫酸铵更适宜于木糖和葡萄糖发酵产乙醇。在摇瓶振荡发酵条件下,该酵母可发酵164.0 g/L葡萄糖生成61.9 g/L乙醇,糖利用率和乙醇得率分别为99.8%和74.0%;受酵母细胞膜上转运体系的限制,对木糖的最高发酵浓度为120.0 g/L,可生成45.7 g/L乙醇,糖利用率和乙醇得率分别达到94.8%和87.0%。休哈塔假丝酵母发酵木糖的主要产物为乙醇,仅生成微量的木糖醇;添加葡萄糖可促进木糖的利用;休哈塔假丝酵母在葡萄糖发酵时的乙酸和甲酸的耐受浓度分别为8.32和2.55 g/L,木糖发酵时的乙酸和甲酸的耐受浓度分别为6.28和1.15 g/L。     

产D-乳酸重组大肠杆菌ptsG基因的敲除及其混合糖同步发酵

为构建能够同时高效利用五碳糖和六碳糖发酵产D-乳酸的重组大肠杆菌工程菌,以能高效利用五碳糖发酵产D-乳酸的大肠杆菌工程菌E.coli JH13为出发菌株,通过Red同源重组技术敲除葡萄糖跨膜转运基因pts G。实验结果表明,pts G缺陷菌株E.coli JH15在10%混合糖(5%葡萄糖和5%木糖)培养基中发酵,可同时利用五碳糖和六碳糖以完成发酵;而对照菌葡萄糖消耗完才利用木糖,发酵结束还有18 g/L木糖残留;JH15乳酸产量为83.04 g/L,相比于对照菌株提高了25.86%;在稻草秸秆水解液中发酵,JH15同时利用葡萄糖、木糖和L-阿拉伯糖,乳酸产量为25.15 g/L,转化率为86.42%。JH15作为能利用混合糖同步发酵产D-乳酸的大肠杆菌工程菌,它的成功构建为利用廉价的木质纤维素水解物为原料发酵生产D-乳酸提供参考依据。     

离子注入选育高产L-乳酸的米根霉突变株及其发酵特性研究

通过氮离子注入获得米根霉突变株RQ4012,其利用木糖的能力比出发菌株提高了1.6倍;通过多次传代,证明其具有良好的遗传稳定性。试验测定菌株RQ4012发酵木糖生产L-乳酸的最佳发酵条件:木糖10%,生理盐水浸泡孢子9 h,(NH4)2SO43 g/L,接种量4%,CaCO3添加量6%,装液量20%,温度37℃,转速200 r/min,在此条件下,乳酸产量达到79.51 g/L。对混合糖的发酵进行了探索,结果表明该菌能高效利用混合糖生产L-乳酸,在利用植物纤维素水解液生产L-乳酸上有良好的应用前景。     

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.