融合株SPSC发酵生产酒精的工艺研究 Ⅰ.絮凝速率、发酵过程的最适温度和pH值

酿酒酵母属(S. cereviae)变异株和粟酒裂殖酵母属(S. pombe)变异株进行属间原生质体融合得到融合株SPSC,该融合株比S. cereviae具有强的自身絮凝能力。以葡萄糖浓度150g/L的底物在30～44℃的温度范围内进行摇瓶厌氧发酵,获得最佳温度范围为34～38℃,最高发酵温度为40℃。在有效容积2.35L悬浮床反应器中,在pH值3.0～5.0范围内进行连续发酵,获得最适发酵pH为3.5～4.5。     

TRIM21 suppresses CHK1 activation by preferentially targeting CLASPIN for K63-linked ubiquitination

Expression of the E3 ligase TRIM21 is increased in a broad spectrum of cancers; however, the functionally relevant molecular pathway targeted by TRIM21 overexpression remains largely unknown. Here, we show that TRIM21 directly interacts with and ubiquitinates CLASPIN, a mediator for ATR-dependent CHK1 activation. TRIM21-mediated K63-linked ubiquitination of CLASPIN counteracts the K6-linked ubiquitination of CLASPIN which is essential for its interaction with TIPIN and subsequent chromatin loading. We further show that overexpression of TRIM21, but not a TRIM21 catalytically inactive mutant, compromises CHK1 activation, leading to replication fork instability and tumorigenesis. Our findings demonstrate that TRIM21 suppresses CHK1 activation by preferentially targeting CLASPIN for K63-linked ubiquitination, providing a potential target for cancer therapy.     

肿瘤浸润淋巴细胞在实体肿瘤中作用的研究进展

肿瘤是21世纪威胁人类健康的主要疾患之一。临床上,实体瘤治疗仍以手术切除、放化疗和靶向治疗为主,但这些方法往往不能根除肿瘤病灶,易导致肿瘤复发和进展。肿瘤免疫治疗是利用人体的免疫系统,通过增强或恢复抗肿瘤免疫力实现控制和杀伤肿瘤的一种新的治疗模式。肿瘤免疫治疗能够在众多患者中产生持久反应,过继性免疫治疗和免疫检查点阻断剂治疗均可产生显著的抗原特异性免疫反应。肿瘤浸润淋巴细胞(TILs)是一种存在于肿瘤组织内部具有高度异质性的淋巴细胞,在宿主抗原特异性肿瘤免疫应答中发挥关键作用。最新研究表明,在肿瘤发生和治疗过程中,TILs的亚群组成和数量与患者预后密切相关;抗肿瘤的TILs介导的过继性免疫治疗方法已在多种实体瘤中取得了良好的疗效。文中就实体肿瘤中TILs的研究进展作一综述。     

Hydrogen exchange: the modern legacy of Linderstrøm-Lang.

This discussion, prepared for the Protein Society's symposium honoring the 100th anniversary of Kaj Linderstrøm-Lang, shows how hydrogen exchange approaches initially conceived and implemented by Lang and his colleagues some 50 years ago are contributing to current progress in structural biology. Examples are chosen from the active protein folding field. Hydrogen exchange methods now make it possible to define the structure of protein folding intermediates in various contexts: as tenuous molten globule forms at equilibrium under destabilizing conditions, in kinetic intermediates that exist for less than one second, and as infinitesimally populated excited state forms under native conditions. More generally, similar methods now find broad application in studies of protein structure, energetics, and interactions. This article considers the rise of these capabilities from their inception at the Carlsberg Labs to their contemporary role as a significant tool of modern structural biology.     

Occurrence and dominance of yeast species in naturally fermented milk from the Tibetan Plateau of China

To determine which yeasts are present in the naturally fermented milks of China, 69 samples made by the nomads of Tibet were collected from the Tibetan Plateau in China. From these samples, 225 strains of yeast were isolated and identified using conventional microbiological analysis and gene sequencing analysis of the D1/D2 domain of the large subunit (26S) ribosomal DNA. The results showed that the total concentration of yeasts in these samples ranged from 5.01 to 8.97 log10 colony-forming units (CFU)/mL (6.91?± 1.02 log10 CFU/mL; mean?± SD). The number of cultivable yeasts was higher in the samples from Qinghai (7.55?± 0.75 log10 CFU/mL) than those from Tibet (6.21?± 0.79 log10 CFU/mL, P?< 0.05). Moreover, there were 15 phylotypes in these 69 samples. Among these phylotypes, Kluyveromyces marxianus (49.3%, frequency percentage), Saccharomyces cerevisiae (62.3%), and Pichia fermentans (46.4%) appeared frequently and can be considered the most common culturable species in naturally fermented milk products. Traditional fermented Mongolian cow milk featured a wide diversity of yeast species, including Issatchenkia orientalis, Kazachstania unisporus, Rhodotorula mucilaginosa, Candida pararugosa, Torulaspora delbrueckii, Geotrichum sp., Kazachstania unisporus, Geotrichum fragrans, Debaryomyces hansenii, Yarrowia lipolytica, Trichosporon gracile, and Pichia membranifaciens. This study provides new data on yeast composition in naturally fermented milk and shows the yeast biodiversity of fermented milk products from the Tibetan Plateau of China.     

一株新的α-淀粉酶抑制剂生产菌株ZG0656及其产物的发酵、分离、性质与应用

从土壤中分离并筛选得到了一株α-淀粉酶抑制剂生产菌,编号ZG0656.根据形态特征,培养特征、生理生化特征、细胞壁化学组成特征和16S rDNA全序列相似性比较分析等多相分类方法,确认菌株ZG0656为天蓝黄链霉菌的新变种,命名为天蓝黄链霉菌南开变种(Streptomyces coelicoflavus var.nankaiensis).该菌经10 L发酵罐水平发酵,发酵液中可积累一定量的α-淀粉酶抑制荆.采用浓缩,树脂吸附,凝胶过滤,减压干燥等方法得到α-淀粉酶抑制剂混合物.该α-淀粉酶抑制剂为含氮的拟低聚糖类物质.能强烈抑制哺乳动物来源的α-淀粉酶,对餐后高血糖的形成有明显改善作用.可用于制备治疗糖尿病,肥胖症的药物或功能性食品.     

26S rDNA单链构象多态性分析在临床酵母菌菌种鉴定中的应用

摘要： 【目的】探讨酵母菌临床分离株26S rDNA D1/D2区序列种内相似性和种间差异性的快速检测方法,为临床酵母菌菌种鉴定方法的改进奠定基础。调查北京地区临床酵母菌的种群多样性,为国内酵母菌感染的流行病学研究提供新的基础数据。【方法】用5种常见临床酵母菌种的模式和权威菌株作为标准参考菌株,从北京四家综合性医院收集临床酵母菌260余株,PCR扩增其26S rDNA D1/D2区,对扩增产物进行单链构象多态性（Single-Strand Conformation Polymorphism,SSCP）分析和序列测定分析。【结果】常见病原酵母菌26S rDNA D1/D2区的SSCP图谱具有明显的种间差异性和种内相似性,可以通过该方法对菌株进行初步的菌种鉴定。D1/D2-SSCP和序列分析相结合,对260余株临床酵母菌进行了菌种鉴定,共鉴定有10个属20个种,优势属为念珠菌属(Candida),优势种及其所占比例分别是：C. albicans (57.7%), C. parapsilosis (10.0%), C. tropicalis (9.2%), C. glabrata (6.7%)和C. krusei (5.8%),并发现过去从未或很少报道致病的酵母菌种,愈来愈多地出现在临床分离菌株中。【结论】 26S rDNA D1/D2区的SSCP图谱分析为临床酵母菌株的快速鉴定提供了新的方法;北京地区酵母菌临床分离株呈种群多样性分布,C. albicans虽然仍占优势,但其它念珠菌种的比例已达42%。     

口蹄疫病毒非结构蛋白3AB双抗体夹心ELISA方法的建立

抗原纯净度是口蹄疫(Foot-and-mouthdisease,FMD)灭活疫苗质量检验的一项重要内容,一般采用疫苗2–3次免疫动物后,检测非结构蛋白(Non-structuralprotein,NSP)抗体是否阳转,判断疫苗抗原的纯净度。文中旨在建立定量检测FMD灭活疫苗抗原中NSP3AB含量的ELISA方法,为疫苗质量控制提供参考方法。利用口蹄疫病毒(Foot-and-mouthdiseasevirus,FMDV)NSP3A单克隆抗体和辣根过氧化物酶(Horseradish peroxidase,HRP)标记的3B单克隆抗体,建立定量检测NSP3AB含量的双抗体夹心ELISA检测方法。采用原核表达并纯化的3AB蛋白作为标准品,标准品系列稀释,绘制标准曲线,以标准品与未加抗原的阴性对照吸光值(OD)的比值大于2.0的标准品最低浓度为最低检测限。标准品浓度介于4.7–600.0 ng/mL之间时,测得的OD值与浓度呈线性相关,回归曲线呈直线,相关系数R2=0.99,确定最低检测限为4.7ng/mL。检测12份未纯化灭活抗原中3AB蛋白含量介于9.3–200.0ng/mL之间;而纯化后的...     

Length–weight relationships for four endemic fish species from the Yalu River,China

Length–weight relationships (LWRs) are presented for four endemic fish species caught in the upper Yalu River during the yearly surveys (2017) by either electrofishing or fish cage net (5 mm mesh size) as soon as the ice melted. The parameter b ranged between 2.244 in Gobio macrocephalus and 3.428 in Ladislavia taczanowskii. Additionally, the parameter a ranged between 0.004 in Misgurnus bipartitus and 0.059 in Gobio macrocephalus. All length‐weight relationships were highly significant (r² ≥ 0.8754) on a log scale.