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Genetic diversity of multidrug resistant Salmonella enterica subsp. enterica serovar Brancaster isolated from chicken in Malaysia 多重耐药肠沙门氏菌亚种的遗传多样性。从马来西亚的鸡中分离出血清型Brancaster肠杆菌
Pub Date : 2023-11-13 DOI: 10.35118/apjmbb.2023.031.4.02
Evie Khoo, Roseliza Roslee, Zunita Zakaria, Nur Indah Ahmad
Salmonella enterica subsp. enterica serovar Brancaster is a chicken-associated serovar that is increasingly reported in the Asian region, including Malaysia. The shortfall of conventional serovar identification for nomenclature alone is insufficient for studying the genetic and clonal relationships of Salmonella of the same serovar. Whereas DNA sequence-based typing method is useful to study the genetic diversity and relatedness of the highly diverse Salmonella. In this study, the genetic diversity, phenotypic antimicrobial resistance (AMR) profiles, and detection of selected AMR genes from ten representative Salmonella Brancaster isolates from chicken cloacal swabs and raw chicken meat were carried out. Based on multilocus sequence typing (MLST), all Salmonella Brancaster belonged to sequence type ST2133. Further analysis of the MLST sequencing data using concatenated nucleotide sequences of the seven housekeeping genes divided them into nine clusters, revealed heterogeneity and genetic diversity among the isolates. All isolates were multidrug-resistant (MDR), which confers resistance to three or more classes of antibiotics. Seven Salmonella Brancaster isolates exhibited phenotypic ACSSuT-type MDR profile, and two of them were also resistant towards ciprofloxacin. Salmonella Brancaster in the present study possessed at least one and up to six resistance genes, with the most prevalent being the ampicillin (blaTEM), tetracycline (tetA), chloramphenicol (floR), gentamicin (ant(3”)-Ia), streptomycin (strA), and sulfonamides (sul-2). Chickens are common reservoir for serovar Brancaster and the emergence, persistence, and dissemination of MDR Salmonella Brancaster possessed a public health risk. The authorities need to strengthen salmonellosis control management programme in poultry farms, abattoirs, and processing facilities.
肠道沙门氏菌亚种Brancaster是一种与鸡相关的血清型肠炎,在包括马来西亚在内的亚洲地区越来越多地报道。传统的血清型命名鉴定的不足不足以研究同一血清型沙门氏菌的遗传和克隆关系。而基于DNA序列的分型方法可用于研究高度多样化的沙门氏菌的遗传多样性和亲缘性。本研究对10株具有代表性的Brancaster沙门氏菌进行了遗传多样性、表型耐药(AMR)分析,并对部分AMR基因进行了检测。基于多位点序列分型(MLST),所有Brancaster沙门氏菌均属于ST2133序列型。利用7个管家基因的核苷酸序列对MLST测序数据进行进一步分析,将它们划分为9个簇,揭示了分离物之间的异质性和遗传多样性。所有分离株均具有多重耐药(MDR),即对三种或更多种抗生素具有耐药性。7株Brancaster沙门氏菌表现出acssut型耐多药表型,其中2株对环丙沙星也有耐药性。本研究中的Brancaster沙门氏菌具有至少一种至六种耐药基因,其中最常见的是氨苄西林(blaTEM)、四环素(tetA)、氯霉素(floR)、庆大霉素(ant(3”)-Ia)、链霉素(strA)和磺胺类药物(su -2)。鸡是血清型布兰卡斯特沙门氏菌的常见宿主,耐多药布兰卡斯特沙门氏菌的出现、持续和传播具有公共卫生风险。当局需要加强家禽养殖场、屠宰场和加工设施的沙门氏菌病控制管理规划。
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引用次数: 0
Novel single nucleotide polymorphism (rs1600485907) of IL-41 gene associated with systemic lupus erythematous IL-41基因与系统性红斑狼疮相关的新单核苷酸多态性(rs1600485907)
Pub Date : 2023-10-22 DOI: 10.35118/apjmbb.2023.031.4.01
Reema Mohammed Abed, Laith Ahmed Yaaqoob
This study evaluated the serum concentrations of IL-41 and the detection of specific single nucleotide polymorphisms (SNPs) (rs1600485907, rs762398841, and rs575275512) within the IL-41 gene in female subjects diagnosed with systemic lupus erythematosus (SLE). The study sample comprised 124 female patients who had been diagnosed with SLE, with an equivalent number of healthy control volunteers. The levels of serum were determined using the application of an enzyme-linked immunosorbent assay (ELISA) technology, while SNPs were determined by conventional polymerase chain reaction (PCR), and subsequent Sanger sequencing. The results revealed that the mean age of the healthy control group was 31.64 years, whereas the mean age of the group diagnosed with SLE was 33.66 years. The findings of the present investigation indicate that the individuals under examination demonstrated an average disease duration of 9.0 years, whereas the average SLEDAI-2k score for those diagnosed with SLE was 11.0. The concentrations of ESR, CRP, urea, creatinine, C3, and C4 in individuals diagnosed with SLE showed a statistically significant elevation in comparison to the control cohort. The analysis of patients’ anti-nuclear antibodies (ANA) revealed that 89% of individuals have ANA, whereas 95.61% display anti-dsDNA. Significantly elevated levels of serum IL-41 were seen in patients diagnosed with SLE in comparison to healthy controls. A receiver operating characteristic (ROC) analysis was performed on a cohort of patients diagnosed with SLE to assess the diagnostic efficacy of IL-41 in discriminating between SLE patients and non-afflicted persons. The study determined that the specificity of IL-41 was 82.26%, representing the percentage of accurate negative outcomes. In contrast, the sensitivity of IL-41 was found to be 84.68%, indicating the percentage of accurate positive outcomes. The computed value for the area under the curve (AUC) was found to be 0.937. The statistical analysis revealed a significant connection between the existence of both heterozygote and homozygote mutant genotypes of IL-41 (rs1600485907) and an increased vulnerability to the formation of SLE. In conclusion, it has been shown that individuals diagnosed with SLE demonstrate heightened concentrations of IL-41in their circulating blood plasma. Furthermore, a particular genetic variation, specifically a single nucleotide polymorphism (SNP) denoted as rs1600485907 located within the IL-41 gene, has been recognized as a potential susceptibility factor for the onset of this disorder.
本研究评估了诊断为系统性红斑狼疮(SLE)的女性受试者血清IL-41浓度和IL-41基因特异性单核苷酸多态性(snp) (rs1600485907、rs762398841和rss575275512)的检测。研究样本包括124名被诊断为SLE的女性患者,以及同等数量的健康对照志愿者。血清水平采用酶联免疫吸附试验(ELISA)技术测定,而单核苷酸多态性采用常规聚合酶链反应(PCR)测定,随后进行Sanger测序。结果显示,健康对照组的平均年龄为31.64岁,SLE诊断组的平均年龄为33.66岁。本研究结果表明,接受检查的个体平均病程为9.0年,而SLE患者的平均SLEDAI-2k评分为11.0。诊断为SLE的个体ESR、CRP、尿素、肌酐、C3和C4的浓度与对照组相比有统计学意义的升高。患者抗核抗体(ANA)分析显示89%的个体具有ANA,而95.61%的个体显示抗dsdna。与健康对照相比,SLE患者血清IL-41水平显著升高。对一组SLE患者进行受试者工作特征(ROC)分析,以评估IL-41在区分SLE患者和非SLE患者中的诊断功效。研究确定IL-41的特异性为82.26%,代表准确的阴性结果百分比。相比之下,IL-41的敏感性为84.68%,表明准确阳性结果的百分比。曲线下面积(AUC)的计算值为0.937。统计分析显示IL-41 (rs1600485907)的杂合子和纯合子突变基因型的存在与SLE易感性的增加之间存在显著的联系。总之,已经证明被诊断为SLE的个体在其循环血浆中il -41浓度升高。此外,一种特殊的遗传变异,特别是位于IL-41基因内的单核苷酸多态性(SNP) rs1600485907,已被认为是该疾病发病的潜在易感因素。
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引用次数: 0
In silico expression profiling and function prediction of transcribed small open reading frames from Cucumis sativus var. hardwickii PI183967 in C. sativus var. sativus sativus var. hardwickii PI183967转录小开放阅读框在sativus var. sativus中的表达分析及功能预测
Pub Date : 2023-10-14 DOI: 10.35118/apjmbb.2023.031.3.10
Gabrielle Shiao Wei Chieng, Boon Chin Tan, Chee How Teo
Cucumis sativus is one of the most cultivated and consumed plants worldwide. To meet the demanding requirements following the rise in population, it is important to maintain and enhance the growth and development of this crop. Recent findings suggest that small open reading frames (sORFs) are associated not only with plant growth but with plant stress responses, especially in the case of drought and salt stress conditions. Hence, in this present study, we aim to determine the function and expression profile reflected in Cucumis sativus var. hardwickii PI183967 sORFs in the variety C. sativus var. sativus through transcriptomic analyses of publicly available RNA-seq datasets of var. sativus. We managed to obtain 14,799 transcribed sORF from the transcriptome datasets of var. sativus. Among these, only 689 (4.66%) transcribed sORF have translational potential. We were able to unveil the roles of transcribed sORFs associated with biological processes (reproductive process, immune response, and multicellular organism reproduction), molecular functions (transferase activity, catalytic activity, acting on a protein, hydrolase activity, and oxidoreductase activity), and cellular component (cytoplasm). KEGG pathway analysis revealed that the transcribed sORFs were significantly enriched in two of the KEGG Ontology (KO) terms, namely, plant-pathogen interaction and plant hormone signal transduction. Results from this study aid the understanding of sORF roles in Cucumis sativus and pave the way for future attempts to utilize these sORFs involved in stress responses to enhance the quality of the crop.
黄瓜是世界上栽培和消费最多的植物之一。为了满足人口增长的需求,保持和提高这种作物的生长和发展是很重要的。最近的研究表明,小开放阅读框(sorf)不仅与植物生长有关,而且与植物的胁迫反应有关,特别是在干旱和盐胁迫条件下。因此,本研究旨在通过公开的sativus var. hardwickii PI183967 sORFs转录组学分析,确定sativus var. hardwickii PI183967 sORFs在sativus var. sativus中的功能和表达谱。我们成功地从sativus变种的转录组数据集中获得了14,799个转录的sORF。其中,只有689个(4.66%)转录的sORF具有翻译潜力。我们能够揭示转录的sorf与生物过程(生殖过程、免疫反应和多细胞生物繁殖)、分子功能(转移酶活性、催化活性、作用于蛋白质、水解酶活性和氧化还原酶活性)和细胞成分(细胞质)相关的作用。KEGG通路分析显示,转录的sorf在KEGG本体(KO)的两个术语,即植物-病原体相互作用和植物激素信号转导中显著富集。本研究的结果有助于理解sORF在黄瓜中的作用,并为今后利用这些sORF参与胁迫应答以提高作物品质铺平道路。
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引用次数: 0
Effects of nitrogen starvation on TAG biosynthesis genes expression in Chlorella vulgaris 氮饥饿对小球藻TAG生物合成基因表达的影响
Pub Date : 2023-09-16 DOI: 10.35118/apjmbb.2023.031.3.08
Jesreen Tega, Cha Thye San, Malinna Jusoh
Microalgal oils and lipids are the potential sources of sustainable industrial products for pharmaceuticals, nutraceuticals, and cosmeceuticals. However, the production cost of microalgal-based products is still expensive and hinders their marketability. Therefore, research has been focusing on increasing microalgae's oil and lipids content to be economically reasonable. Nutrient stresses were often used to enhance oils and lipids production in microalgae. In this study, the microalga Chlorella vulgaris was exposed to nitrogen starvation for 21 days to examine the effect of nitrogen removal on cell density, total oil content, fatty acids composition, and the expression of two triacylglycerols (TAG) biosynthetic genes, glycerol-3-phosphate acyltransferase (GPAT) and diacylglycerol acyltransferase (DGAT). The results showed that the C. vulgaris cells grew extremely slowly (p<0.05) under nitrogen starvation conditions. However, despite the growth outcome, the nitrogen starvation increased (p<0.05) total oil contents on days 3, 11 and 14 relatives to control. Nitrogen starvation also stimulated (p<0.05) the production of saturated fatty acids (SFA). The primary fatty acids detected were C16:0, C18:0, C18:1, C18:2, C18:3 and C20:0. The effect of nitrogen starvation on the expression of TAG biosynthetic genes, GPAT and DGAT genes were enumerated using real-time PCR. Both GPAT and DGAT were downregulated in this study. Interestingly, the buildup of C18:2 and C18:3 was positively linked with GPAT expression, demonstrating that GPAT affected the synthesis of polyunsaturated fatty acids (PUFA) in nitrogen-starved circumstances. This suggests that the GPAT gene may be altered to increase PUFA in microalgae, notably C18:2 and C18:3.
微藻油和脂质是可持续工业产品的潜在来源,如药品、营养保健品和药妆品。然而,基于微藻的产品的生产成本仍然昂贵,阻碍了它们的市场化。因此,提高微藻油脂含量以达到经济合理的目的一直是研究的重点。营养胁迫常被用来提高微藻的油脂产量。本研究以普通小球藻(Chlorella vulgaris)为试验材料,对其进行了21 d的氮饥饿处理,研究脱氮对细胞密度、总油含量、脂肪酸组成以及甘油-3-磷酸酰基转移酶(GPAT)和二酰基甘油酰基转移酶(DGAT)两种三酰基甘油(TAG)生物合成基因表达的影响。结果表明,在氮饥饿条件下,C. vulgaris细胞生长极慢(p<0.05)。然而,与对照组相比,氮饥饿在第3、11和14天使总油含量增加(p<0.05)。氮饥饿也刺激了饱和脂肪酸(SFA)的产生(p<0.05)。检测到的主要脂肪酸为C16:0、C18:0、C18:1、C18:2、C18:3和C20:0。采用实时荧光定量PCR技术,分析氮饥饿对TAG生物合成基因、GPAT和DGAT基因表达的影响。GPAT和DGAT在本研究中均下调。有趣的是,C18:2和C18:3的积累与GPAT的表达呈正相关,表明GPAT影响了氮饥饿环境下多不饱和脂肪酸(PUFA)的合成。这表明GPAT基因可能被改变以增加微藻中的PUFA,特别是C18:2和C18:3。
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引用次数: 0
Characterization of fumarate reduction by Klebsiella pneumoniae isolated from patients with chronic periodontitis 慢性牙周炎患者分离的肺炎克雷伯菌对富马酸还原的研究
Pub Date : 2023-09-16 DOI: 10.35118/apjmbb.2023.031.3.07
Geetika Sharma, Nancy Garg, Shamimul Hasan, Daad Saffarini, Sheetal Shirodkar
Chronic periodontitis is oral inflammatory gum disease which affects the tissues that support the teeth. It is caused by formation of bacterial plaque consisting of bacterial species that produces various virulence factors. Oral bacterial species from the periodontitis patients include species of Fusobacterium, Porphyromonas, Prevotella, Tannerella, etc. In the present study, we isolated the Klebsiella pneumoniae strain 9A from the pockets of patients suffering from chronic periodontitis. The strain was isolated on enriched tryptic soya media containing 10 mM fumarate and identified based on Gram staining, 16S rRNA gene sequencing, biochemical tests and ability to reduce fumarate. Our study is the first report of anaerobic fumarate reduction capabilities of Klebsiella spp. isolated from chronic periodontitis patients, which may help in growth and survival of species in periodontal pockets leading to progression of the disease. In addition, experimental evidence is provided through reduction, growth, and inhibitor assays for the presence of fumarate reductase (Frd) activities. Furthermore, we detected a 400 bp frdA gene fragment in Klebsiella pneumoniae strain 9A using degenerate PCR. This fragment show similarity to fumarate reductases flavoprotein subunit (FrdA). Our findings suggest that the FrdA subunit is responsible for anaerobic fumarate respiration in Klebsiella spp. to be important in the survival of bacteria in oxygen-deprived environments in humans such as periodontal pockets, thus contributing to its pathogenicity.
慢性牙周炎是一种口腔炎症性牙龈疾病,它会影响支撑牙齿的组织。它是由细菌菌斑的形成引起的,菌斑由产生各种毒力因子的细菌种类组成。牙周炎患者口腔细菌种类包括梭杆菌、卟啉单胞菌、普雷沃菌、坦纳菌等。在本研究中,我们从慢性牙周炎患者的口袋中分离出肺炎克雷伯菌9A菌株。菌株在含有10 mM富富富马酸的胰蛋白酶大豆培养基上分离,通过革兰氏染色、16S rRNA基因测序、生化试验和还原富马酸的能力对菌株进行鉴定。我们的研究首次报道了从慢性牙周炎患者中分离的克雷伯氏菌的厌氧富马酸还原能力,这可能有助于牙周袋中物种的生长和生存,从而导致疾病的进展。此外,通过还原、生长和抑制剂试验提供了富马酸还原酶(Frd)活性存在的实验证据。此外,我们还利用退化PCR技术在肺炎克雷伯菌9A株中检测到一个400 bp的frdA基因片段。该片段与富马酸还原酶黄蛋白亚基(FrdA)相似。我们的研究结果表明,FrdA亚基负责克雷伯氏菌的厌氧富马酸呼吸,对细菌在人类缺氧环境(如牙周袋)中的生存至关重要,从而有助于其致病性。
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引用次数: 0
Review of bioactive components property of Malaysian propolis: A review 马来西亚蜂胶生物活性成分性质研究进展
Pub Date : 2023-09-16 DOI: 10.35118/apjmbb.2023.031.3.09
Khor Goot Heah, Eleena Mohd Yusof, Sharvitha Dhamotharan, Ikmal Hisham Ismail
Over the past few decades, traditional medicinal products have grown in popularity. It is estimated that 80% of patients worldwide use them for basic health treatment. Due to its extraordinary bioactive properties, propolis is a bee product commonly utilised in alternative medicines. Propolis is a complicated mixture of substances, procured by bees from diverse vegetations and mix them with beeswax and bee saliva enzymes. Propolis bioactivities have been examined and reported in the treatments of numerous chronic diseases. Given the differences in pharmacological activity and compound markup of propolis, huge variation exists due to numerous hive localities. This study intends to define and explore the bioactive qualities of propolis in Malaysia. Information on the antibacterial, antioxidant, anti-inflammatory, and other bioactivity properties from studies conducted on Malaysian propolis have been consolidated in this review. This research would result in an enhanced understanding of the safe consumption of Malaysian propolis and may spur the development of novel and more affordable treatments for various infections.
在过去的几十年里,传统医药产品越来越受欢迎。据估计,全世界80%的患者使用它们进行基本卫生治疗。由于其非凡的生物活性特性,蜂胶是一种常用的替代药物的蜂产品。蜂胶是一种复杂的物质混合物,由蜜蜂从不同的植被中获取,并将其与蜂蜡和蜜蜂唾液酶混合。蜂胶的生物活性已在许多慢性疾病的治疗中得到检验和报道。鉴于蜂胶的药理活性和化合物标记的差异,由于蜂房位置众多,蜂胶存在巨大的差异。本研究旨在定义和探索马来西亚蜂胶的生物活性特性。关于马来西亚蜂胶的抗菌、抗氧化、抗炎和其他生物活性特性的研究已经在这篇综述中得到了巩固。这项研究将使人们对马来西亚蜂胶的安全消费有更深入的了解,并可能促进开发新的、更实惠的治疗各种感染的方法。
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引用次数: 0
Effect of salinity and biopesticide on Arthrospira platensis mass culture growth, metabolites, and bacterial diversity 盐度和生物农药对platarthrospira mass culture生长、代谢物和细菌多样性的影响
Pub Date : 2023-09-02 DOI: 10.35118/apjmbb.2023.031.3.06
Irma Rohmawati, B. K. Adji, Dea Putri Andeska, E. Suyono
Arthrospira platensis mass culture contamination is a recurring concern. Salinity alteration appears to be a promising approach, given that A. platensis can withstand various salinities. In addition, biopesticide is also expected to eliminate the bacteria. Therefore, it is critical to investigate the effects of salinity and biopesticide on the growth, nutritional value, and bacterial diversity of A. platensis mass culture. A completely randomized design was used, with five salinity treatment levels: 5 ppt (S5B), 10 ppt (S10B), 15 ppt (S15B), 20 ppt (S20B), and 25 ppt (S25B), with 0.5 ml/L of biopesticide Azadirachta indica applied on days 2 and 4. The growth rate was calculated using daily density and biomass, and on day 7, carbohydrates, proteins, lipids, and pigments were determined. Furthermore, the contamination test and bacterial diversity were determined using Total Plate Count and Next-Generation Sequencing, respectively. The results showed that the S15B had the fastest growth rate and the largest carbohydrate content. However, S5B produced the best results in terms of protein, chlorophyll, and phycocyanin content, while S25B produced the most carotene and lipids. Proteobacteria were the most abundant in all NGS samples. The number of OTU treatments obtained for A (NCD7), B (NCD0), and C (S15BD7) were 646, 636, and 286, respectively. Moreover, C (S15BD7) was the most effective treatment to reduce bacterial diversity contamination, in which several bacterial diversity, including Acidobacteriota, Chloroflexi, Gemmatimonadota, Myxococcota, and Desulfobacterota, were eliminated completely. As a result of the salinity adjustment and the use of biopesticides, the mass culture of A. platensis was able to grow more quickly and contain more nutrients while having less bacterial diversity.
platarthrospira mass culture污染是一个反复出现的问题。盐度变化似乎是一种很有前途的方法,因为platensis可以承受各种盐度。此外,生物农药也有望消除细菌。因此,研究盐度和生物农药对白颡鱼生长、营养价值和细菌多样性的影响具有重要意义。采用完全随机设计,5个盐度处理水平分别为5 ppt (S5B)、10 ppt (S10B)、15 ppt (S15B)、20 ppt (S20B)和25 ppt (S25B),在第2天和第4天分别施用0.5 ml/L的印楝生物农药。利用日密度和生物量计算生长速率,并在第7天测定碳水化合物、蛋白质、脂质和色素。此外,污染测试和细菌多样性分别使用Total Plate Count和下一代测序进行测定。结果表明,S15B的生长速度最快,碳水化合物含量最高。然而,在蛋白质、叶绿素和藻蓝蛋白含量方面,S5B产生的效果最好,而S25B产生的胡萝卜素和脂质最多。变形菌门在所有NGS样品中数量最多。A (NCD7)、B (NCD0)和C (S15BD7)获得的OTU处理数分别为646、636和286。此外,C (S15BD7)是减少细菌多样性污染最有效的处理,可以完全消除酸菌群、氯氟菌群、双胞菌群、粘菌群和脱硫菌群等细菌多样性。由于盐度的调整和生物农药的使用,大量培养的platensis能够更快地生长,含有更多的营养,但细菌多样性较少。
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引用次数: 0
Preliminary molecular identification of proteolytic andlipolytic-enzyme producing bacteria isolated from sediment of Litopenaeus vannamei pond 凡纳滨对虾池塘沉积物中蛋白水解和脂酶产菌的初步分子鉴定
Pub Date : 2023-09-02 DOI: 10.35118/apjmbb.2023.031.3.05
D. Ayuningrum, D. Novitasari, A. Sabdaningsih, O. Jati
The enzyme is a catalyst that serves as an accelerator in a biochemical process. A hydrolytic enzyme is among the existing enzymes useful for environmental organic waste degradation. This research aimed to (1) investigate the ability of bacteria isolated from Litopenaeus vannamei pond sediment to produce proteolytic and lipolytic enzymes, (2) identify the isolates with 16S rRNA gene amplification, and (3) construct a phylogenetic tree according to the 16S rRNA genes. The preliminary proteolytic assay consisted of a skim milk agar medium and the lipolytic assay consisted of a Tween 20/80 medium or so-called precipitation test. Among 28 bacterial isolates, 4 of them showed potential for proteolytic activity and 6 of them showed lipolytic activity as well. Further, the amplification of 16S rRNA gene showed that 5 out of the 28 isolates were closely related to Bacillus infantis SA 3.2 (IM8), Marinobacter koreensis SB 1.1 (IM6), Vibrio algynolyticus SA 4.2 (IM6), Streptomyces euryhalinus SB 1.2 (IM6), and Cytobacillus kochii SC 3.4 (IM6). Amongst the most active isolate, one of the was included in Actinobacteria phylum. Many Bacilli strains shows enzymatic activity such as amylase, protease and lypase. The genus Marinobacter also found to be able to produce hydrolytic enzyme such as amylase and protease. Furthermore, the genus Vibrio such as V. algynoliticus produce several enzymes i.e., gelatinase, lecithinase, caseinase, amylase and lipase. In summary, the bacteria from L. vannamei pond sediment exhibit a potential as proteolytic and lipolytic enzyme producers.
这种酶是一种催化剂,在生化过程中起到加速器的作用。水解酶是对环境有机废物降解有用的现有酶之一。本研究旨在(1)研究凡纳滨对虾池塘沉积物中分离的细菌产生蛋白水解酶和脂肪水解酶的能力,(2)鉴定具有16S rRNA基因扩增的分离菌株,(3)根据16S rRNA基因构建系统发育树。初步的蛋白水解试验由脱脂牛奶琼脂培养基组成,脂溶试验由Tween 20/80培养基或所谓的沉淀试验组成。28株分离菌株中,4株具有潜在的蛋白水解活性,6株具有潜在的脂肪水解活性。通过16S rRNA基因扩增,28株分离菌中有5株与婴儿芽孢杆菌SA 3.2 (IM8)、朝鲜海洋杆菌SB 1.1 (IM6)、藻解弧菌SA 4.2 (IM6)、泛盐链霉菌SB 1.2 (IM6)、kochi胞杆菌SC 3.4 (IM6)亲缘关系较近。在最活跃的分离物中,其中一个被列入放线菌门。许多芽孢杆菌菌株具有淀粉酶、蛋白酶和酶解酶等酶活性。海洋杆菌属也被发现能够产生水解酶,如淀粉酶和蛋白酶。此外,弧菌属如藻酸弧菌可产生多种酶,即明胶酶、卵磷脂酶、酪蛋白酶、淀粉酶和脂肪酶。综上所述,凡纳梅l池塘沉积物中的细菌显示出作为蛋白质水解酶和脂肪水解酶生产者的潜力。
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引用次数: 0
In silico analysis of xylanase in Bacillus coagulans ST-6 凝固芽孢杆菌ST-6中木聚糖酶的硅化分析
Pub Date : 2023-08-27 DOI: 10.35118/apjmbb.2023.031.3.03
A. M. Suhaimi, Rabiatul Adawiah Zainal Abidin, Fairuz Fatini Mohd Yusof, Abdullah Munir Roslan, A. Sipat, K. Yusoff
Bacillus coagulans ST-6 is a thermophile isolated from a local hot spring in Malaysia. It expresses xylanase activity and has potential industrial applications. In silico structure prediction and modeling of a 210 amino acid sequence (XYNBC) translated from 872 bp Bacillus coagulans ST-6 xylanase gene was performed. The predicted molecular weight of the translated amino acid sequence is 20 kDa with theoretical isoelectric point value at 9.10 predicting that the xylanase belongs to the GH11 xylanase family. The protein is predicted to be hydrophilic with Grand average of hydropathicity (GRAVY) value of -0.6555 and possibly has better interaction with water. Predicted motif for XYNBC was revealed to be from Glycosyl hydrolases family 11(IPR00137) with signature motif 1 and 2 and only one known activity, xylanase. A three-dimensional (3D) model was constructed using PDB ID 2DCZ|A (http://www.rcsb. org) as the template as it has the highest similarity with XYNBC protein sequence. The 2DCZ|A xylanase sequence is from Bacillus Subtilis family-11 and is 185 amino acid long. The predicted 3D model consists of eleven beta sheets and one alpha helix. The stereochemical quality of protein structure revealed by Ramachandran Plot showed acceptable model with 99.4% residues fall in the most favored regions. Ten residues were predicted to be involved in active sites where residues Tyr7, Val15 and Asn16 are located at beta sheet while Asp118, Gly119, Thr120 are located at the loop. The catalytic residues E78 and E172 common to other G1H1 xylanases were also revealed.
凝结芽孢杆菌ST-6是一种从马来西亚当地温泉分离出来的嗜热菌。它表达木聚糖酶活性,具有潜在的工业应用价值。对从872 bp的凝固芽孢杆菌ST-6木聚糖酶基因中翻译的210个氨基酸序列(XYNBC)进行了硅结构预测和建模。预测翻译氨基酸序列的分子量为20 kDa,理论等电点值为9.10,预测该木聚糖酶属于GH11木聚糖酶家族。预测该蛋白具有亲水性,亲水性(GRAVY)的大平均值为-0.6555,可能与水有较好的相互作用。预测的XYNBC基序来自糖基水解酶家族11(IPR00137),其特征基序为1和2,只有一个已知的活性,即木聚糖酶。使用PDB ID 2DCZ|A (http://www.rcsb)构建三维(3D)模型。org)作为模板,因为它与XYNBC蛋白序列相似性最高。2DCZ|A木聚糖酶序列来自枯草芽孢杆菌家族11,全长185个氨基酸。预测的3D模型由11个β层和一个α螺旋组成。Ramachandran图显示的蛋白质结构立体化学质量符合可接受的模型,99.4%的残基落在最有利的区域。预计有10个残基参与活性位点,其中残基Tyr7、Val15和Asn16位于β片上,而残基Asp118、Gly119、Thr120位于环上。同时还发现了其他G1H1木聚糖酶共有的催化残基E78和E172。
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引用次数: 0
Phytochemical and cytotoxic properties of Solanum aethiopicum fruit extracts against HeLa human cervical cancer cell line 埃塞俄比亚茄果实提取物对HeLa人宫颈癌细胞系的植物化学和细胞毒性研究
Pub Date : 2023-08-27 DOI: 10.35118/apjmbb.2023.031.3.04
Lim Yee Thing, Lejaniya Abdul Kalam Saleena, Crystale Lim Siew Ying, R. Koh, Pui Liew Phing
Eggplant, also known as Solanum aethiopicum (S. aethiopicum), is a highly favoured vegetable globally and belongs to the Solanaceae family. Various research studies have provided evidence of African eggplant's biological properties, such as antioxidant potential, anti-inflammatory, anti-cancer, anti-diabetic, liver-protective, and renal-protective effects. Therefore, our investigation focused on examining the impact of S. aethiopicum on the cytotoxicity of HeLa cells, which are derived from human cervical cancer. Fruits were freeze-dried or oven-dried before being subjected to aqueous infusion or hot aqueous extraction. Hot aqueous extracted (oven-dried) samples contained the highest concentration of total phenol and flavonoids. After that, HeLa cells were treated with hot aqueous extracts. After a 24-hour incubation period, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was conducted. According to the cytotoxicity analysis, neither oven-dried nor freeze-dried induced significant inhibitory effects towards HeLa cell proliferation. These findings suggest that both extracts demonstrated weak cytotoxic effects on cervical cancer cells.
茄子,也被称为埃塞俄比亚茄(S. aethiopicum),是一种全球备受青睐的蔬菜,属于茄科。各种研究已经为非洲茄子的生物学特性提供了证据,如抗氧化潜力、抗炎、抗癌、抗糖尿病、保护肝脏和保护肾脏的作用。因此,我们的研究重点是研究埃塞俄比亚梭菌对人宫颈癌HeLa细胞的细胞毒性的影响。水果被冷冻干燥或烘箱干燥,然后再进行水浸或热水萃取。热水萃取(烘箱干燥)样品含有最高浓度的总酚和黄酮类化合物。之后,用热水提取物处理HeLa细胞。24小时孵育后,进行3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑(MTT)试验。根据细胞毒性分析,干燥和冷冻干燥对HeLa细胞增殖均没有明显的抑制作用。这些发现表明,这两种提取物对宫颈癌细胞的细胞毒性作用较弱。
{"title":"Phytochemical and cytotoxic properties of Solanum aethiopicum fruit extracts against HeLa human cervical cancer cell line","authors":"Lim Yee Thing, Lejaniya Abdul Kalam Saleena, Crystale Lim Siew Ying, R. Koh, Pui Liew Phing","doi":"10.35118/apjmbb.2023.031.3.04","DOIUrl":"https://doi.org/10.35118/apjmbb.2023.031.3.04","url":null,"abstract":"Eggplant, also known as Solanum aethiopicum (S. aethiopicum), is a highly favoured vegetable globally and belongs to the Solanaceae family. Various research studies have provided evidence of African eggplant's biological properties, such as antioxidant potential, anti-inflammatory, anti-cancer, anti-diabetic, liver-protective, and renal-protective effects. Therefore, our investigation focused on examining the impact of S. aethiopicum on the cytotoxicity of HeLa cells, which are derived from human cervical cancer. Fruits were freeze-dried or oven-dried before being subjected to aqueous infusion or hot aqueous extraction. Hot aqueous extracted (oven-dried) samples contained the highest concentration of total phenol and flavonoids. After that, HeLa cells were treated with hot aqueous extracts. After a 24-hour incubation period, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was conducted. According to the cytotoxicity analysis, neither oven-dried nor freeze-dried induced significant inhibitory effects towards HeLa cell proliferation. These findings suggest that both extracts demonstrated weak cytotoxic effects on cervical cancer cells.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79959834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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Asia-pacific Journal of Molecular Biology and Biotechnology
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