产几丁质酶菌株SWCH-6的筛选、鉴定及其产酶条件的优化研究

从汕头海湾养殖区域的海底沉积物中分离到1株几丁质酶活性较高的菌株,命名为SWCH-6,根据菌株的形态特征、生理生化特征和16S Rdna序列,确定该菌株为嗜水气单胞茵(Aeromonas hydrophlilla).采用单因素优化方法结合正交实验,得到菌株SWCH-6产几丁质酶的最佳发酵条件:胶体几丁质25.0g/L,胰蛋白胨10.0g/L,陈海水1.0L, Ph 8.5,32℃,150 r/min培养72h;在该条件下酶活力达0.39U/Ml.此外,菌株所产几丁质酶的最适催化Ph 5.0;最适催化温度为40℃;Cu2、Fe3及表面活性剂Tween-80能增强该酶的催化活性;Zn2 、Mn2 及表面活性剂SDS、洗衣粉对该酶的催化活性有抑制作用,与其它几丁质酶存在着一些不同.     

莱氏野村菌产几丁质酶条件及酶学性质研究

对莱氏野村菌(Nomuraea rileyi)菌株CQ031021产几丁质酶条件及酶学性质进行了研究。结果表明:该菌株最适产酶碳源为2.0%(W/V)葡萄糖,氮源为1.2%(W/V)复合氮源(蛋白胨、牛肉膏按1∶1的比例),接种量为孢悬液2mL(1×107个/mL),培养温度28℃,培养液初始pH6.0,培养时间6d;一定浓度的吐温-80对几丁质酶活性有促进作用,而SDS有抑制作用;粗酶液最适反应温度50℃,最适pH6.0,在40℃以下及pH5.5～6.5范围内酶活力较稳定。     

假交替单胞菌LJ1菌株产褐藻胶裂解酶的培养条件优化及酶学性质

[目的]为了优化Lj1菌株的培养条件使之产生高活性的胞外褐藻胶裂解酶.[方法]通过富集培养技术从海带筛选到一株褐藻胶裂解酶产生菌Lj1,依据表型特征、脂肪酸组成分析及16S rRNA基因序列分析对该菌株进行鉴定.通过单因子和正交试验对Lj1菌株产胞外褐藻胶裂解酶的培养条件进行了优化.[结果]Lj1菌株属于假交替单胞菌属(Pseudoalteromonas).该菌株产酶的最佳培养基组成为:褐藻胶3g/L、(NH4)2SO43 g/L、NaCl 20 g/L、KH2PO4 0.1 g/L、CaCl2 0.1 g/L;最佳培养条件为:250 mL三角烧瓶中装液量25 mL、接种量3%、摇瓶转速150 r/min、pH7.5、培养温度为28℃、培养时间为24 h.LJl菌株所产褐藻胶裂解酶的最适温度为40℃,最适pH7.6,最适NaCl浓度为0.3 mol/L.1 mol/1.金属离子Mg2+对酶活力有明显的促进作用,而C02+和Zn2+对酶活力有较强的抑制作用.[结论]LJ1菌株是Pseudoalteromonas新的胞外褐藻胶裂解酶产生菌,在最佳培养条件下,该菌株的酶活力提高了66%.     

煤附生真菌产漆酶菌株的分离鉴定及产酶特性研究

从煤炭样品中筛选到一株产漆酶活性菌株,经菌体形态观察和ITS序列分析,鉴定为Trichoderma asperellum W03。菌株所产漆酶的最适反应pH为3.5-4.5,最适反应温度45℃,类似于白腐真菌漆酶。液态发酵条件的均匀设计实验表明,适宜的发酵培养基组成为:土豆200.00g/L、葡萄糖9.36g/L、米糠粉37.44g/L、硝酸钾4.00g/L、KH2PO43.20g/L、MgSO4·7H2O2.00g/L、CuSO4·5H2O0.005g/L、初始pH8.0;在33℃、180r/min、50mL/250mL的摇瓶培养条件下,棘孢木霉W03在孢子接种培养后48h、84h产酶量较高,分别处在菌体的快速生长期和衰亡期;菌体产酶受Cu2+、联苯胺诱导,而受1-萘酚、愈创木酚和2,4-D抑制。     

一步法发酵菊芋产乙醇菌种的筛选及产酶条件、酶学性质的研究

从腐烂的菊芋及实验室保存的菌种中,选育到一株发酵菊芋产乙醇的菌株克鲁维酵母Kluyveromyces marxianus Y1。利用正交实验法对克鲁维酵母产菊粉酶的培养基组成及培养条件进行优化,确定培养基组成（g/L）为：菊粉40,酵母粉4,蛋白胨4,尿素1;初始pH5．0,温度30℃,150r／min条件下培养达到最佳产酶效果（57U／mL）。该菌株所产菊粉酶的性质测定结果表明：以菊粉为底物,该菊粉酶最适反应温度为55℃,在60℃以下稳定性很好,高于60℃时酶迅速失活;最适pH为5．0,pH4．6—5．2范围内酶稳定性很好;该酶属于外切型菊粉酶,体积分数为8％的乙醇对酶活力基本没有影响。     

产木聚糖酶白地霉培养特性及部分纯化的酶学特性

本文对白地霉Ref1的培养特性、产酶条件和酶学特性进行了初步研究。结果表明:该菌为低温型菌株,其最佳生长条件为pH6、20℃和酵母膏作为氮源;最佳产酶条件为pH3-7、15℃及以酵母膏氮源;条件优化后产酶可达118.7U/mL,可溶蛋白含量可达到60μg/mL,酶溶液的比活可达到1250U/mg蛋白质;该木聚糖酶的最适反应温度和pH分别为50℃和5,金属离子Mg2+、Na+和8mmol/L的Fe2+、Cu2+、Zn2+等对木聚糖酶的活性有抑制作用,而Ca2+、4mmol/L的Fe2+、Cu2+、Zn2+和8mmol/L的Mn2+等对该酶反应则有促进作用;该木聚糖酶在保温2h后在15-40℃范围内能保持80%以上的酶活性,在50℃时能保持68%的酶活性;用lineweaver-Burk作图法(双倒数作图法)求得该酶的最大反应速度Vmax和Km值分别为163.38mmol/mg/min和0.75mg/mL。     

褐藻胶裂解酶产生菌的分离鉴定及产酶发酵优化

对一株从腐烂海带中筛选得到的产褐藻胶裂解酶的菌株进行鉴定,并对其产酶条件进行发酵优化。经形态学、生理生化特征和分子生物学鉴定,将其鉴定为盐单胞菌属,并命名为Halomonas sp. WF6。通过在摇瓶培养水平上进行单因素和多因素正交试验,确定褐藻胶裂解酶产生菌WF6的最适产酶培养基为：褐藻酸钠6.0 g/L,蛋白胨5.0 g/L,酵母粉2.5 g/L,NaCl 30 g/L,K⁺ 5 mmol/L。进而采用最适培养基进行产酶条件的优化,优化后的发酵产酶条件为：初始pH 8.0,培养温度25℃,接种量为2%,摇瓶装液量30 ml/250 ml,培养时间39 h。优化后的褐藻胶裂解酶酶活达117.66 U/ml,是优化前的2.1倍。该酶对褐藻酸钠的酶解产物主要由聚合度为二和三的褐藻寡糖组成。     

高温蛋白酶产生菌的筛选及其产酶条件和酶学性质分析

从徂徕山温泉附近土样中分离到9株产高温蛋白酶菌株,选取一株碱性蛋白酶高产菌株L7为出发菌株,进行显微形态、16S rRNA基因序列分析,将其初步鉴定为短芽孢杆菌(Brevibacillus sp.)。研究该菌株发酵条件,确定产酶的最佳培养基组成为葡萄糖40 g/L,蛋白胨20 g/L,磷酸氢二钠1.4 g/L,氯化钙0.6 g/L,硫酸镁0.4 g/L,通过培养基优化,酶活达到103.08 U/mL。最佳培养条件为250 mL三角烧瓶中装液量50 mL、pH8.0、培养温度为55℃、培养时间为24 h。对该菌株酶学性质研究,L7菌株所产高温蛋白酶的最适温度为55℃,最适pH为10,并且具有良好的温度稳定性和pH稳定性,酶活性受PMSF强烈抑制。     

绿僵菌Ma83几丁质酶的发酵研究

本实验从虫生真菌中筛选出金龟子绿僵菌M a83菌株,它的几丁质酶合成能力最强。其产酶的适宜条件是,碳源为胶体几丁质加葡萄糖,氮源为NaNO3,培养温度为28℃,培养基起始pH 6.0;接种量为5 mL液态种,最适装液量为5 mL,添加维生素C可以提高酶活;正交实验表明培养因子的最佳组合是:NaNO31 g/L,胶体几丁质0.6 g/L,酵母膏0.05 g/L,葡萄糖0.10 g/L。根据液态培养产酶过程结果可知,当M a83菌培养6天时,几丁质酶活力达到8.1 U/mL。     

几丁质酶产生菌筛选鉴定及产酶性能研究

从土壤样品中筛选得到一株高产几丁质酶菌株C65-2,经形态学观察和18S rDNA序列测定,鉴定为Aspergillus fumigatus,对产酶培养基进行初步优化,测得最高酶活可达6.9U/ml,酶活力较优化之前提高了210%。酶学性质研究表明该几丁质酶分子量约为20kDa,酶在60℃下保温50min酶活降为0,最适酶反应温度是55℃,酶反应最适pH为7.0,Mg2+,Cu2+对酶反应有促进作用,Fe3+对酶反应有抑制作用。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.