元阳豆豉中高产蛋白酶乳酸菌的筛选及其产酶条件的研究

从云南省元阳地区采集豆豉样品,并从中分离得到62株乳酸菌菌株。通过脱脂乳平板试验,从中初步筛得到21株具有高蛋白酶活力的菌株,采用茚三酮法测定其蛋白酶活力,复筛出高产蛋白酶菌株YY-1-6L,并对其产酶条件进行研究。结果表明,YY-1-6L在以葡萄糖为碳源、多聚蛋白胨为氮源、起始pH 5.0的明胶诱导培养基中,接种量为3%,35℃发酵培养36 h,其蛋白酶活力高达32.50 U/mL,且K2HPO4、MgSO4能促进蛋白酶产生。     

Characterization of a novel mutant KCNQ1 channel subunit lacking a large part of the C-terminal domain

A mutation of KCNQ1 gene encoding the alpha subunit of the channel mediating the slow delayed rectifier K⁺ current in cardiomyocytes may cause severe arrhythmic disorders. We identified KCNQ1(Y461X), a novel mutant gene encoding KCNQ1 subunit whose C-terminal domain is truncated at tyrosine 461 from a man with a mild QT interval prolongation. We made whole-cell voltage-clamp recordings from HEK-293T cells transfected with either of wild-type KCNQ1 [KCNQ1(WT)], KCNQ1(Y461X), or their mixture plus KCNE1 auxiliary subunit gene. The KCNQ1(Y461X)-transfected cells showed no delayed rectifying current. The cells transfected with both KCNQ1(WT) and KCNQ1(Y461X) showed the delayed rectifying current that is thought to be mediated largely by homomeric channel consisting of KCNQ1(WT) subunit because its voltage-dependence of activation, activation rate, and deactivation rate were similar to the current in the KCNQ1(WT)-transfected cells. The immunoblots of HEK-293T cell-derived lysates showed that KCNQ1(Y461X) subunit cannot form channel tetramers by itself or with KCNQ1(WT) subunit. Moreover, immunocytochemical analysis in HEK-293T cells showed that the surface expression level of KCNQ1(Y461X) subunit was very low with or without KCNQ1(WT) subunit. These findings suggest that the massive loss of the C-terminal domain of KCNQ1 subunit impairs the assembly, trafficking, and function of the mutant subunit-containing channels, whereas the mutant subunit does not interfere with the functional expression of the homomeric wild-type channel. Therefore, the homozygous but not heterozygous inheritance of KCNQ1(Y461X) might cause major arrhythmic disorders. This study provides a new insight into the structure–function relation of KCNQ1 channel and treatments of cardiac channelopathies.     

Isolation,molecular characterization and antibiotic resistance of Shiga Toxin–Producing Escherichia coli (STEC) from buffalo in India

In total, 363 Escherichia coli were isolated from 165 faecal samples of healthy buffaloes in West Bengal, India. Twenty‐four of these isolates (6·61%) were found to carry at least one gene characteristic for Shiga toxin–producing Escherichia coli (STEC). These STEC strains belonged to 13 different O‐serogroups. The stx₁ gene was present in 23 (95·8%) of total STEC isolates, whereas 20 (83·3%) STEC isolates carried the gene stx_2. Twelve strains of E. coli (50% of total STEC isolates) possessed enterohaemolysin (ehxA) gene in combination with others. Fourteen (58·33%) isolates found to possess saa gene. However, no E. coli was detected harbouring gene for intimin protein (eaeA). Of 23 stx₁‐positive isolates, seven (30·43%) were positive for genes of the stx_1C subtype. Of the 20 isolates with the stx₂ gene, 25% (5/20) possessed stx_2C and 10% (2/20) possessed stx_2d gene. The phylogenetic analysis after RAPD of STEC strains revealed six major clusters. The isolated STEC strains were resistant most frequently to erythromycin (95·83%), cephalothin (62·5%), amikacin (54·17%), kanamycin (45·83%) and gentamicin (41·67%) group of antibiotics. No ESBL‐producing (bla_CTXM, bla_TEM, bla_SHV) or quinolone resistance gene (qnrA) was detected in the STEC isolates.

Significance and Impact of the Study

The buffaloes from different districts of West Bengal, India, are important reservoir of multidrug‐resistant Shiga toxin–producing Escherichia coli (STEC). India is home to more than 56% of world buffalo population, traditionally raised by farmers. So, there is a major risk of transmission of STEC among the human population of this part of the globe. However, there is no prevalence study of STEC from healthy or diarrhoeic buffalo in India. The present study reports for the first time in India about isolation, molecular characterization and antibiotic resistance pattern of STEC in healthy buffaloes.     

高产不饱和油脂海鞘真菌桔青霉Asc-2-4的诱变选育

诱变育种是获得高产菌株,实现微生物工业化生产油脂的重要措施。以前期获得的高产不饱和油脂菌株桔青霉（Penicillium citrinum）Asc-2-4为出发菌株,利用丙二酸建立快速筛选高产不饱和脂肪酸突变菌的方法,通过紫外线 氯化锂复合诱变得到1株高产油脂突变菌Asc-2-4-1,油脂含量比出发菌株提高了92.98%。经过初步的培养基无机盐优化,其油脂得率和不饱和脂肪酸产量达到了7.10 g/L和3.84 g/L,与Asc-2-4相比,分别提高了84.42%和77.78%。结果表明,通过复合诱变选育技术可选育出高产突变菌株,选育的Asc-2-4-1可望作为产油微生物被开发利用。     

CHLOROPHYLL C PIGMENT PATTERNS IN 18 SPECIES (51 STRAINS) OF THE GENUS PSEUDO‐NITZSCHIA (BACILLARIOPHYCEAE)1

The pigment composition of 18 species (51 strains) of the pennate diatom Pseudo‐nitzschia was examined using HPLC. The carotenoid composition was typical for diatoms, with fucoxanthin (the major xanthophyll), diadinoxanthin, diatoxanthin, and β,β‐carotene. However, a diverse array of chl c pigments was observed in the studied strains. All Pseudo‐nitzschia strains contained chl a and chl c₂, traces of Mg‐2,4‐divinyl phaeoporphyrin a₅ monomethyl ester (MgDVP), and traces of a chl c₂–like pigment originally found in the haptophyte Pavlova gyrans. The distribution of chl c₁ and chl c₃ was variable among species (present in seven and 14 species, respectively). Based on chl c distribution, three major pigment types were defined: type 1 (chl c₁ + c₂, four species: P. australis, P. brasiliana, P. multiseries, and P. seriata), type 2 (chl c₁ + c₂ + c₃, three species: P. fraudulenta, P. multistriata, and P. pungens), and type 3 (chl c₂ + c₃, 11 species: P. arenysensis, P. calliantha, P. cuspidata, P. decipiens, P. delicatissima, P. galaxiae, P. mannii, P. pseudodelicatissima, P. subcurvata, P. cf. subpacifica, and a novel Pseudo‐nitzschia species). Type 1 and 2 species also shared the absence of a particular morphological character, the central nodule in the raphe, with the only exception of P. fraudulenta. The implications of such pigment diversity in chemotaxonomy, HAB monitoring, ecology, and phylogeny of Pseudo‐nitzschia species are discussed.     

Phylogenomics and multigene phylogenies decipher two new cryptic marine algae from California,Gelidium gabrielsonii and G. kathyanniae (Gelidiales,Rhodophyta)

Molecular surveys are leading to the discovery of many new cryptic species of marine algae. This is particularly true for red algal intertidal species, which exhibit a high degree of morphological convergence. DNA sequencing of recent collections of Gelidium along the coast of California, USA, identified two morphologically similar entities that differed in DNA sequence from existing species. To characterize the two new species of Gelidium and to determine their evolutionary relationships to other known taxa, phylogenomic, multigene analyses, and morphological observations were performed. Three complete mitogenomes and five plastid genomes were deciphered, including those from the new species candidates and the type materials of two closely related congeners. The mitogenomes contained 45 genes and had similar lengths (24,963–24,964 bp). The plastid genomes contained 232 genes and were roughly similar in size (175,499–177,099 bp). The organellar genomes showed a high level of gene synteny. The two Gelidium species are diminutive, turf‐forming, and superficially resemble several long established species from the Pacific Ocean. The phylogenomic analysis, multigene phylogeny, and morphological evidence confirms the recognition and naming of two new species, describe herein as G. gabrielsonii and G. kathyanniae. On the basis of the monophyly of G. coulteri, G. gabrielsonii, G. galapagense, and G. kathyanniae, we suggest that this lineage likely evolved in California. Organellar genomes provide a powerful tool for discovering cryptic intertidal species and they continue to improve our understanding of the evolutionary biology of red algae and the systematics of the Gelidiales.     

The iterative process of plant species inventorying for obtaining reliable biodiversity patterns

We require representative data of species occurrence to explain plant diversity patterns, but most of the available information is incomplete and biased. To improve our knowledge, we suggest that species inventorying should be an iterative process encompassing the following: (1) the detection of taxonomic and geographical gaps; (2) the planning of a survey design to reduce such gaps; and (3) the evaluation of field sampling results. Here, we focus on the latter phase for the bryophytes of Terceira Island (Azores) for which we have previously estimated < 1% of the area as well surveyed based on historical collections. To examine the performance of our stratified survey based on two factors (land use and environmental regions), we used rarefaction curves, ANOVA tests and bootstrap sampling. We recorded 40% of all the species known for the island and presented eight new citations. The species assemblages remained similar between historical and current inventories. Most localities had completeness values > 85%, but we always exceeded the optimal sampling effort. Land uses and environmental regions affected species diversity, but, unexpectedly, to a different degree. Our study illustrates the difficulties of planning field surveys to obtain reliable biodiversity patterns, even when prior information and standardized sampling protocols are explicitly considered. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 491–503.     

A chalcone with potent inhibiting activity against biofilm formation by nontypeable Haemophilus influenzae

Nontypeable Haemophilus influenzae (NTHi), an important human respiratory pathogen, frequently causes biofilm infections. Currently, resistance of bacteria within the biofilm to conventional antimicrobials poses a major obstacle to effective medical treatment on a global scale. Novel agents that are effective against NTHi biofilm are therefore urgently required. In this study, a series of natural and synthetic chalcones with various chemical substituents were evaluated in vitro for their antibiofilm activities against strong biofilm‐forming strains of NTHi. Of the test chalcones, 3‐hydroxychalcone (chalcone 8 ) exhibited the most potent inhibitory activity, its mean minimum biofilm inhibitory concentration (MBIC₅₀) being 16 μg/mL (71.35 μM), or approximately sixfold more active than the reference drug, azithromycin (MBIC₅₀ 419.68 μM). The inhibitory activity of chalcone 8 , which is a chemically modified chalcone, appeared to be superior to those of the natural chalcones tested. Significantly, chalcone 8 inhibited biofilm formation by all studied NTHi strains, indicating that the antibiofilm activities of this compound occur across multiple strong‐biofilm forming NTHi isolates of different clinical origins. According to antimicrobial and growth curve assays, chalcone 8 at concentrations that decreased biofilm formation did not affect growth of NTHi, suggesting the biofilm inhibitory effect of chalcone 8 is non‐antimicrobial. In terms of structure–activity relationship, the possible substituent on the chalcone backbone required for antibiofilm activity is discussed. These findings indicate that 3‐hydroxychalcone (chalcone 8 ) has powerful antibiofilm activity and suggest the potential application of chalcone 8 as a new therapeutic agent for control of NTHi biofilm‐associated infections.     

12种不同产地铁皮石斛指纹图谱研究及重金属元素含量测定

以12种不同产地铁皮石斛新鲜茎段为材料,以Ultimate XB C18(4.6mm×250mm,5μm)乙腈-0.1%磷酸溶液为流动相进行梯度洗脱,流速1.0mL·min-1,检测波长270nm,柱温30℃。通过中药色谱指纹图谱相似度评价系统(国家药典委员会)进行相似度分析,并建立12个不同产地铁皮石斛指纹图谱。经微波消解处理材料后,采用ICP-AES原子吸收光谱法测定不同产地铁皮石斛新鲜茎段中重金属铜、镉、铅、汞的含量,分析不同产地铁皮石斛质量,为建立铁皮石斛质量标准提供理论依据。结果表明:(1)不同产地铁皮石斛指纹图谱共有23个特征峰,相似度为0.918~0.987,其中产自安徽大别山、云南文山的铁皮石斛相似度分别为0.918、0.935,相比其他产地具有一定的差异性。(2)通过聚类分析将12种不同产地铁皮石斛分为4类:安徽霍山、江苏南京、云南石屏、云南玉溪、云南思茅为一类,云南文山、浙江仙居为一类,云南红河、广西天峨、安徽大别山、福建宁化为一类,浙江天台单独为一类。(3)产自云南玉溪的铁皮石斛茎段中铜含量超标26.25%,产自福建宁化和江苏南京的镉含量分别超标27.33%和122.30%,其它产地铜、镉、铅、汞含量在国家现行安全值以内。