Optimization of L-asparaginase production from Escherichia coli using response surface methodology

Among the antitumor drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. In previous study, the L-asparaginase from Erwinia chrysanthermy was expressed in Escherichia coli BL21(DE3). The recombinant L-asparaginase was produced from recombinant E.coli BL21(DE3) under different cultivation conditions (inducer concentration, inoculum concentration and KH2PO4 concentration). The optimized conditions by response surface methodology using face centered central composite design. The analysis of variance coupled with larger value of R2 (0.9) showed that the quadratic model used for the prediction was highly significant (p 0.05). Under the optimized conditions, the model produced L-asparaginase activity of 123.74 U/ml at 1.03 mM IPTG, 3% (v/v) inoculum and 0.5% (w/v) KH2PO4. Recombinant protein was purified by two step using gel filtration and DEAE chromatography. The purified L-asparaginase had a molecular mass of 37 kDa with specific activity of 462 U/mg and identified by MALDI-TOF mass spectrometry. Results of MALDI-TOF analysis confirmed that recombinant protein was L-asparaginase II. Recombinant L-asparaginase has antiproliferative activity with K562 cell line. In conclusion, this study has innovatively developed cultivation conditions for better production of recombinant L-asparaginase in shake flask culture

Phrasal semantic distance for vietnamese textual document retrieval

In this paper, a computational semantic method is proposed to estimate the phrasal semantic distance used in our model of a Vietnamese document retrieval system. The semantic distances between phrases are defined in terms of semantic classes and semantic relations to ensure that it can reflect how different two certain phrases are. To estimate the semantic distance, the semantic classes of a phase are identified by using the n-gram model. After identification of the semantic classes, their semantic relations are also identified by using a Vietnamese Lexicon Ontology. This handcrafted ontology contains defined semantic classes and their potential relations in Vietnamese language explicitly. For the evaluation purpose, a phrasal semantic retrieval system has been built to test with a data set of 720 phrases and 30 queries. The evaluation shows the precision of 96.6% and the recall of 78.4% on experimental results

Mutagenesis development of actinoplanes sp. KCTC 9161 by N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and screening for acarbose production

Acarbose has been widely used in the therapy of type II diabetes (non-insulin dependent) because it controls blood sugar contents of patients after meals. Acarbose, a pseudo-oligosaccharide, acts as a competitive -glucosidase inhibitor. Acarbose is produced by the strains of Bacillus, Streptomyces and Actinoplanes sp. The aim of this study was to develop mutagenesis for an Actinoplanes sp. strain and screening for acarbose production. The spores of Actinoplanes sp. KCTC 9161 strain were subjected to be mutated by N-methyl-N'-nitro-N-nitrosoguanidine (NTG) for screening and finding mutant strains that were capable of production of higher acarbose (an inhibitor of α-glucosidase) higher than wild type strain. Firstly, the original NTG solution was prepared in phosphate buffer 0.05 M, pH 6.9 and the safety concentration of NTG was determined at 5 mg/ml. Then, the spores were incubated with different NTG amounts and duration. The living colonies were transferred to fermentation medium. The results obtained showed that 15 mutant strains were produced higher acarbose than wild type when used thin layer chromatography method for analysis and comparing with standard acarbose (Sigma). Three cell lines among total tested 15 mutant lines of Actinoplanes sp. KCTC 9161 produced acarbose at a higher level or indicated a higher inhibitory activity toward α-glucosidase than the original strain. Enzymatic inhibitory ativity of α-glucosidase of three mutant strains (Actinoplanes sp. KCTC- L4, L11, L14) was increased 1.3 fold higher than wild type and Actinoplanes sp. KCTC spores were very sensitive to NTG toxic, 98% spores could not survive at the treatment condition of 50 µg NTG for 30 minutes. In addition, an applicable protocol for mutating Actinoplanes sp. using NTG was suggested for further research

IDENTIFICATION AND CHARACTERIZATION OF A PURPLE NONSULFUR BACTERIUM ISOLATED FROM COASTAL AREA OF HAI PHONG FOR USING IN PRODUCTION OF UNSATURATED FATTY ACID (OMEGA 6, 7, 9)

Purple nonsulfur bacteria are a group that has so much biotechnological applications, particularly in producing of functional food rich with unsaturated fatty acids. A purple nonsulfur bacterium (named HPB.6) was chosen based on its strong growth, high lipid and synthesis of unsaturated fatty acid (omega 6,7,9). Studying on basic biological characteristics showed that the cells of HPB.6 were observed as ovoid-rod shape, none motility, Gram negative staining. The diameter of single bacterium was about 0.8-1.0 µm. The cells divide by binary fission and had bacteriochlorophyll a (Bchl a). This bacterium grew well on medium with carbon and nitrogen sources such as acetate, succinate, pyruvate, butyrate, glutamate, arginine, leucine, tyrosine, alanine, methionine, threonine, glutamine, yeast extract and NH4Cl. This selected strain grew well on medium with salt concentrations from 1.5 - 6.0% (optimum 3%), pH from 5.0 to 8.0 (optimum at pH 6.5) and could withstand Na2S at 4.0 - 5.2 mM. Based on morphological, physiological properties and 16S rRNA analysis received demonstrated that HPB.6 strain belongs to the species Rhodovulum sulfidophilum

Using metagenomics tool to evaluate the enrichment efficiency of methanogens in marine sediment in Truong Sa archipelage, Khanh Hoa province, Vietnam

Methanogens (archaea bacteria that produce methan) play an important role in the anaerobic digestion of organic waste, however, in marine environment, the low density of methanogens makes anaerobic digestion very slow. In this study, we used methagenomics tool to evaluate the enrichment efficiency of methanogens communities in marine sediments as a basis for building a anaerobic microbial formulation to treat organic waste in marine environment. The results of determination of methanogen density by MPN method have shown that methanogen has been accumulated with higher density (up to 3.2.107 MNP/ml) through 2 times of enrichment in artificial seawater with CH3COONa 10 mM substrate. In addition, metagenomics data have also shown a decrease in the number of archaea species through each enrichment, indicating that methanogenic species have gradually dominated the microbial community. This is the first study on using metagnenomics tools to evaluate the enrichment of methanogens in marine sediment samples in Truong Sa archipelago, Vietnam. Metagenomics data help provide more reliable evidence in microbial studies, especially in obligate anaerobes such as methanogens

Excess body weight and age associated with the carriage of fluoroquinolone and third-generation cephalosporin resistance genes in commensal Escherichia coli from a cohort of urban Vietnamese children.

PURPOSE: Antimicrobial-resistant bacterial infections in low- and middle-income countries (LMICs) are a well-established global health issue. We aimed to assess the prevalence of and epidemiological factors associated with the carriage of ciprofloxacin- and ceftriaxone-resistant Escherichia coli and associated resistance genes in a cohort of 498 healthy children residing in urban Vietnam. METHODOLOGY: We cultured rectal swabs onto MacConkey agar supplemented with resistant concentrations of ciprofloxacin and ceftriaxone. Additionally, we screened meta-E. coli populations by conventional PCR to detect plasmid-mediated quinolone resistance (PMQR)- and extended-spectrum β-lactamase (ESBL)-encoding genes. We measured the associations between phenotypic/genotypic resistance and demographic characteristics using logistic regression.Results/Key findings. Ciprofloxacin- and ceftriaxone-resistant E. coli were cultured from the faecal samples of 67.7 % (337/498) and 80.3 % (400/498) of children, respectively. The prevalence of any associated resistance marker in the individual samples was 86.7 % (432/498) for PMQR genes and 90.6 % (451/498) for β-lactamase genes. Overweight children were significantly more likely to carry qnr genes than children with lower weight-for-height z-scores [odds ratios (OR): 1.24; 95 % confidence interval (CI): 10.5-1.48 for each unit increase in weight for height; P=0.01]. Additionally, younger children were significantly more likely to carry ESBL CTX-M genes than older children (OR: 0.97, 95 % CI: 0.94-0.99 for each additional year, P=0.01). CONCLUSION: The carriage of genotypic and phenotypic antimicrobial resistance is highly prevalent among E. coli in healthy children in the community in Vietnam. Future investigations on the carriage of antimicrobial resistant organisms in LMICs should focus on the progression of carriage from birth and structure of the microbiome in obesity

No Clinical Benefit of Empirical Antimicrobial Therapy for Pediatric Diarrhea in a High-Usage, High-Resistance Setting.

BACKGROUND: Pediatric diarrheal disease presents a major public health burden in low- to middle-income countries. The clinical benefits of empirical antimicrobial treatment for diarrhea are unclear in settings that lack reliable diagnostics and have high antimicrobial resistance (AMR). METHODS: We conducted a prospective multicenter cross-sectional study of pediatric patients hospitalized with diarrhea containing blood and/or mucus in Ho Chi Minh City, Vietnam. Clinical parameters, including disease outcome and treatment, were measured. Shigella, nontyphoidal Salmonella (NTS), and Campylobacter were isolated from fecal samples, and their antimicrobial susceptibility profiles were determined. Statistical analyses, comprising log-rank tests and accelerated failure time models, were performed to assess the effect of antimicrobials on disease outcome. RESULTS: Among 3166 recruited participants (median age 10 months; interquartile range, 6.5-16.7 months), one-third (1096 of 3166) had bloody diarrhea, and 25% (793 of 3166) were culture positive for Shigella, NTS, or Campylobacter. More than 85% of patients (2697 of 3166) were treated with antimicrobials; fluoroquinolones were the most commonly administered antimicrobials. AMR was highly prevalent among the isolated bacteria, including resistance against fluoroquinolones and third-generation cephalosporins. Antimicrobial treatment and multidrug resistance status of the infecting pathogens were found to have no significant effect on outcome. Antimicrobial treatment was significantly associated with an increase in the duration of hospitalization with particular groups of diarrheal diseases. CONCLUSIONS: In a setting with high antimicrobial usage and high AMR, our results imply a lack of clinical benefit for treating diarrhea with antimicrobials; adequately powered randomized controlled trials are required to assess the role of antimicrobials for diarrhea

Partial food systems baseline assessment at the Vietnam benchmark sites

Using data collected from a cross-sectional study in Moc Chau, Dong Anh and Cau Giay districts in Vietnam, this report aims to elucidate specific components of local Vietnamese food systems along a rural to urban transect focusing specifically on (i) diets, (ii) nutrition status (anthropometry), (iii) consumer behavior, (iv) food environment, and (v) food flows

Genomic serotyping, clinical manifestations, and antimicrobial resistance of non-typhoidal Salmonella gastroenteritis in hospitalized children in Ho Chi Minh City, Vietnam

Nontyphoidal Salmonella (NTS) are among the most common etiological agents of diarrheal diseases worldwide and have become the most commonly detected bacterial pathogen in children hospitalized with diarrhea in Vietnam. Aiming to better understand the epidemiology, serovar distribution, antimicrobial resistance (AMR), and clinical manifestation of NTS gastroenteritis in Vietnam, we conducted a clinical genomics investigation of NTS isolated from diarrheal children admitted to one of three tertiary hospitals in Ho Chi Minh City. Between May 2014 and April 2016, 3,166 children hospitalized with dysentery were recruited into the study; 478 (∼15%) children were found to be infected with NTS by stool culture. Molecular serotyping of the 450 generated genomes identified a diverse collection of serogroups (B, C1, C2 to C3, D1, E1, G, I, K, N, O, and Q); however, Salmonella enterica serovar Typhimurium was the most predominant serovar, accounting for 41.8% (188/450) of NTS isolates. We observed a high prevalence of AMR to first-line treatments recommended by WHO, and more than half (53.8%; 242/450) of NTS isolates were multidrug resistant (MDR; resistant to ≥3 antimicrobial classes). AMR gene detection positively correlated with phenotypic AMR testing, and resistance to empirical antimicrobials was associated with a significantly longer hospitalization (0.91 days; P = 0.04). Our work shows that genome sequencing is a powerful epidemiological tool to characterize the serovar diversity and AMR profiles in NTS. We propose a revaluation of empirical antimicrobials for dysenteric diarrhea and endorse the use of whole-genome sequencing for sustained surveillance of NTS internationally