Maltogenic amylase is one of the significant enzymes in oligosaccharides synthesis. Its ability to utilise multiple substrates and catalyse hydrolysis and transglycosylation reactions simultaneously makes it a unique biocatalyst. The catalysis could be exploited in many ways to obtain oligosaccharides of different lengths and various modified sugars. Nonetheless, one of the major drawbacks of substrate hydrolysis to produce oligosaccharides is the low production of MOS with higher degree of polymerisation. To address this issue, reaction parameter optimisation was performed via one-factor-at-a-time (OFAT) approach on the production of MOS from soluble starch hydrolysis using maltogenic amylase from Bacillus lehensis G1 (MAG1). Optimisation of MAG1 loading, soluble starch loading, temperature, time and pH resulted in the production of 84.87 mg/g MOS with polymerisation degree of 3 to 7 compared to that of 51.60 mg/g obtained before the optimisation process, which recorded 1.64-fold increment. Among all parameters, soluble starch loading gave the most significant impact on the MOS production as the reaction equilibrium is highly affected by substrate concentration. The occurrence of MOS with polymerisation degree of 4 and above, which resulted from starch hydrolysis further confirms the endo-type of MAG1. Because starch is an abundant and inexpensive source of carbohydrate in the world, this study provides a cost-effective MOS production process which is highly relevant for industry.
Constitutive androstane receptor (CAR) encoded by the nuclear receptor subfamily 1, group I, member 3 (NR1I3) gene regulates the elimination of bilirubin through activating the components of the bilirubin clearance pathway. Hence, NR1I3 genetic variants may affect bilirubin metabolism and result in neonatal hyperbilirubinemia. Thus far, research which investigates the association between NR1I3 variants and neonatal hyperbilirubinemia has not been undertaken in any population. The present study aimed to evaluate the influence of MPJ6_1I3008 (rs10157822), IVS8+116T>G (rs4073054) and 540A>G (rs2307424) on neonatal hyperbilirubinemia development in the Malay population. Buccal swabs were collected from 232 hyperbilirubinemia and 277 control term newborns with gestational age ≥37weeks and birth weight ≥2500g. The NR1I3 variants were genotyped by using high resolution melting (HRM) assays and verified by DNA sequencing. Gender, mode of delivery and birth weight did not differ between hyperbilirubinemia and control groups. The genotypic and allelic frequencies of MPJ6_1I3008, IVS8+116T>G and 540A>G were not significantly different between the groups. However, stratification by gender revealed a significant inverse association between homozygous variant genotype of MPJ6_1I3008 and risk of neonatal hyperbilirubinemia in the females (OR, 0.44; 95% CI, 0.20-0.95; p=0.034). This study demonstrates that the homozygous variant genotype of MPJ6_1I3008 was associated with a significant reduced risk of neonatal hyperbilirubinemia in the females.
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