A comprehensive Thai pharmacogenomics database (TPGxD-1): Phenotype prediction and variants identification in 942 whole-genome sequencing data.

Computational methods analyze genomic data to identify genetic variants linked to drug responses, thereby guiding personalized medicine. This study analyzed 942 whole-genome sequences from the Electricity Generating Authority of Thailand (EGAT) cohort to establish a population-specific pharmacogenomic database (TPGxD-1) in the Thai population. Sentieon (version 201808.08) implemented the GATK best workflow practice for variant calling. We then annotated variant call format (VCF) files using Golden Helix VarSeq 2.5.0 and employed Stargazer v2.0.2 for star allele analysis. The analysis of 63 very important pharmacogenes (VIPGx) reveals 85,566 variants, including 13,532 novel discoveries. Notably, we identified 464 known PGx variants and 275 clinically relevant novel variants. The phenotypic prediction of 15 VIPGx demonstrated a varied metabolic profile for the Thai population. Genes like CYP2C9 (9%), CYP3A5 (45.2%), CYP2B6 (9.4%), NUDT15 (15%), CYP2D6 (47%) and CYP2C19 (43%) showed a high number of intermediate metabolizers; CYP3A5 (41%), and CYP2C19 (9.9%) showed more poor metabolizers. CYP1A2 (52.7%) and CYP2B6 (7.6%) were found to have a higher number of ultra-metabolizers. The functional prediction of the remaining 10 VIPGx genes reveals a high frequency of decreased functional alleles in SULT1A1 (12%), NAT2 (84%), and G6PD (12%). SLCO1B1 reports 20% poor functional alleles, while PTGIS (42%), SLCO1B1 (4%), and TPMT (5.96%) showed increased functional alleles. This study discovered new variants and alleles in the 63 VIPGx genes among the Thai population, offering insights into advancing clinical pharmacogenomics (PGx). However, further validation is needed using other computational and genotyping methods.

Characterization of levansucrase produced by novel Bacillus siamensis and optimization of culture condition for levan biosynthesis.

Levan has attracted interest due to the potential health benefits associated with its prebiotic, biological, and functional properties. However, the production of levan is expensive due to its high resource requirements. With the growing demand for levan, it is vital to determine suitable cultivation condition for its production and reduce costs accordingly. The present study characterized the enzyme levansucrase produced by a novel strain of Bacillus siamensis and optimized the conditions for the biosynthesis of levansucrase and levan. The crude levansucrase enzyme production by B. siamensis was induced at a specific temperature in a medium containing different concentrations of sucrose, fructose, and glucose to evaluate transfructosylation and hydrolysis activities. Crude levansucrase significantly increased transfructosylation relative to hydrolysis activity at 37 °C in a medium containing 20% (w/v) sucrose. Both transfructosylation and hydrolysis activities were inhibited in glucose and fructose containing medium. Purification and characterization of the levansucrase were performed by precipitating the enzyme with ammonium sulfate solution, purified anion-exchange chromatography, and analyzed by Sodium Dodecyl Sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed the molecular weight of the enzyme to be approximately 30 kDa with specific activity at 15.95 U/mg, corresponding to a protein purification efficiency of 11.47 and a yield of 78.75%. The optimal culture condition for the purified-levansucrase activity for levan biosynthesis was obtained at 37 °C after 48 h, at pH 6.0 in 50 mM phosphate buffer and 20% (w/v) sucrose. The study demonstrated the optimized condition for levan biosynthesis utilizing the B. siamensis that can serve as a model for various commercial and industrial applications for efficient levan production.

Composition bias and the origin of ORFan genes.

MOTIVATION: Intriguingly, sequence analysis of genomes reveals that a large number of genes are unique to each organism. The origin of these genes, termed ORFans, is not known. Here, we explore the origin of ORFan genes by defining a simple measure called 'composition bias', based on the deviation of the amino acid composition of a given sequence from the average composition of all proteins of a given genome. RESULTS: For a set of 47 prokaryotic genomes, we show that the amino acid composition bias of real proteins, random 'proteins' (created by using the nucleotide frequencies of each genome) and 'proteins' translated from intergenic regions are distinct. For ORFans, we observed a correlation between their composition bias and their relative evolutionary age. Recent ORFan proteins have compositions more similar to those of random 'proteins', while the compositions of more ancient ORFan proteins are more similar to those of the set of all proteins of the organism. This observation is consistent with an evolutionary scenario wherein ORFan genes emerged and underwent a large number of random mutations and selection, eventually adapting to the composition preference of their organism over time.

Identification of beta-lactamase in antibiotic-resistant Bacillus cereus spores.

Beta-lactamase type I is reported for the first time to occur in the sporulated form in a penicillin-resistant Bacillus species. The enzyme was readily characterized from the B. cereus 5/B line (ATCC 13061) by mass spectrometry and two-dimensional gel electrophoresis.