Inverse Association of Serum Folate Level with Metabolic Syndrome and Its Components in Korean Premenopausal Women: Findings of the 2016-2018 Korean National Health Nutrition Examination Survey.

Research on the association of serum folate levels with metabolic syndrome (MetS) in premenopausal women is lacking. This study was aimed to investigate this association in 1730 premenopausal women using the 2016 to 2018 Korean National Health and Nutrition Examination Survey data. Participants' mean age and BMI were 35.9 years and 22.7 kg/m2, respectively. Participants were divided into three groups according to serum folate tertiles. Odds ratios (OR) and 95% confidence intervals (CIs) for abdominal obesity, elevated blood pressure (BP), high fasting plasma glucose (FPG), high triglycerides (TG), low high-density lipoprotein cholesterol (HDL-C), and MetS were calculated in multiple logistic regression models adjusted for possible confounders, by serum folate level tertiles. Prevalence of MetS (14.9, 11.0, and 8.6%); abdominal obesity (17.8, 16.0, and 11.4%); high TG (17.5, 14.0, and 11.1%); and low HDL-C (50.3, 44.6, and 42.5%) decreased with increasing folate level tertile. Prevalence of elevated BP (14.3, 12.0, and 11.7%) and high FPG (11.9, 15.8, and 13.0%) showed no significant differences according to serum folate level tertiles. The multivariate-adjusted ORs (95% CIs) for MetS, abdominal obesity, elevated BP, high TG, and low HDL-C in the highest folate level tertile were 2.17 (1.46-3.22), 1.80 (1.25-2.60), 1.77 (1.16-2.70), 1.90 (1.35-2.67), and 1.49 (1.14-1.94), respectively. The ORs for high FPG did not show significant differences according to serum folate level tertiles. In conclusion, serum folate levels were inversely associated with an increased risk of MetS in Korean premenopausal women. These results suggest that MetS can be prevented and managed by improving the serum folate levels in premenopausal women.

Development and Application of Cyclodextrin Hydrolyzing Mutant Enzyme Which Hydrolyzes ß- and γ-CD Selectively.

Cyclodextrins (CDs) are produced from starch by cyclodextrin glucanotransferase (CGTase), which has cyclization activity. Specifically, α-CD is an important biomolecule, as it is a molecular carrier and soluble dietary fiber used in the food industry. Upon inspection of the conserved regions of the glycoside hydrolase (GH) 13 family amylases, the amino acids K232 and H233 of CGTase were identified as playing an important role in enzyme reaction specificity. A novel CD hydrolyzing enzyme, cyclodextrin glycosyl transferase (CGTase)-alpha, was developed using site-directed mutagenesis at these positions. Action pattern analysis using various substrates revealed that CGTase-alpha was able to hydrolyze ß- and γ-CD, but not α-CD. This selective CD hydrolyzing property was employed to purify α-CD from a CD mixture solution. The α-CD that remained after treatment with CGTase-alpha and exotype glucoamylase was purified using hydrophobic interaction chromatography with 99% purity.

Characterization and Application of BiLA, a Psychrophilic α-Amylase from Bifidobacterium longum.

In this study, a novel α-amylase was cloned from Bifidobacterium longum and named BiLA. The enzyme exhibited optimal activity at 20 °C and a pH value of 5.0. Kinetic analysis using various carbohydrate substrates revealed that BiLA had the highest k(cat/)K(m) value for amylose. Interestingly, analysis of the enzymatic reaction products demonstrated that BiLA specifically catalyzed the hydrolysis of oligosaccharides and starches up to G5 from the nonreducing ends. To determine whether BiLA can be used to generate slowly digestible starch (SDS), starch was treated with BiLA, and the kinetic parameters were analyzed using porcine pancreatic α-amylase (PPA) and amyloglucosidase (AMG). Compared to normal starch, BiLA-treated starch showed lower k(cat)/K(m) values with PPA and AMG, suggesting that BiLA is a potential candidate for the production of SDS.

Characterization of a Novel Maltose-Forming α-Amylase from Lactobacillus plantarum subsp. plantarum ST-III.

A novel maltose (G2)-forming α-amylase from Lactobacillus plantarum subsp. plantarum ST-III was expressed in Escherichia coli and characterized. Analysis of conserved amino acid sequence alignments showed that L. plantarum maltose-producing α-amylase (LpMA) belongs to glycoside hydrolase family 13. The recombinant enzyme (LpMA) was a novel G2-producing α-amylase. The properties of purified LpMA were investigated following enzyme purification. LpMA exhibited optimal activity at 30 °C and pH 3.0. It produced only G2 from the hydrolysis of various substrates, including maltotriose (G3), maltopentaose (G5), maltosyl ß-cyclodextrin (G2-ß-CD), amylose, amylopectin, and starch. However, LpMA was unable to hydrolyze cyclodextrins. Reaction pattern analysis using 4-nitrophenyl-α-d-maltopentaoside (pNPG5) demonstrated that LpMA hydrolyzed pNPG5 from the nonreducing end, indicating that LpMA is an exotype α-amylase. Kinetic analysis revealed that LpMA had the highest catalytic efficiency (kcat/Km ratio) toward G2-ß-CD. Compared with ß-amylase, a well-known G2-producing enzyme, LpMA produced G2 more efficiently from liquefied corn starch due to its ability to hydrolyze G3.

Development and characterization of cyclodextrin glucanotransferase as a maltoheptaose-producing enzyme using site-directed mutagenesis.

Cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) mainly produces cyclodextrins (CDs) using linear maltooligosaccharides. We performed site-directed saturation mutagenesis on the +1 substrate-binding residue, H233, of CGTase from alkalophilic Bacillus sp. I-5 to prepare specific-length oligosaccharides. The obtained mutant CGTase, H233Y, primarily produced maltoheptaose (G7) using ß-CD via a hydrolysis reaction. A kinetic study of H233Y showed that the kcat/Km value of ß-CD was 7-fold greater than that of G7, which accounts for the accumulation of G7 during the H233Y enzyme reaction. A structure comparison of CGTases with H233Y modeling suggests that the substitution of H233Y may alter the position of the +1 subsite and slow the further hydrolysis of G7 after the ring-opening reaction.