Cytochrome P450 (CYPs) catalyze regioselective oxidation reactions, yet the structural determinants governing regioselectivity and catalytic efficiency remain poorly understood. Here, we characterize CYP154C10 as a versatile steroid hydroxylase exhibiting predominant C2 regioselectivity and expanded substrate scope relative to previously described CYP154C enzymes. Notably, CYP154C10 supports steroid conversion using both chemical (PIDA) and NADPH-dependent (Fdx/FdR) systems, while maintaining consistent product distribution across both oxidation systems. Structure-guided mutagenesis, kinetic analysis, and molecular dynamics (MD) simulations together provide mechanistic insights into the functional separation between active-site and access-channel residues. Mutation of the active-site residue L93 to phenylalanine (L93F) induces substrate-dependent changes in regioselectivity by increasing conformational flexibility and enabling multiple catalytically competent substrate orientations. In contrast, mutations at the access-channel position (M199F/Y) preserve regioselectivity but modulate catalytic efficiency by perturbing substrate positioning and local hydration dynamics. Comparative structural and dynamic analyses further suggest that active-site architecture governs substrate orientation, whereas access-channel residues influence substrate delivery and catalytic efficiency in CYP154C10. Together, these findings provide a mechanistic framework for the rational engineering of selective P450 biocatalysts with tunable regioselectivity and catalytic efficiency.

β-agarase is the key enzyme for the primary degradation of agar, a major component of red algae. This study demonstrates cloning, overexpression and characterization of a recombinant GH16 β-agarase (WP_199597959) from Antarctic bacterium Gelidibacter salicanalis PAMC21136. Biochemical properties demonstrated that the maximum activity was observed at pH and temperature of 7.0 and 50°C, respectively using agarose as a substrate. The agarolytic activity of WP_199597959 on agarose was found to be endo-type with production of neoagarobiose (NA2) as a major product. Mn2+ metal ion significantly enhanced the activity, doubling the reaction rate as compared with other metal ions. Neoagarooligosaccharides (NAOSs) like neoagarotetraose (NA4), and neoagarohexaose (NA6) hydrolyzed into NA2. In-silico analysis demonstrated that the key catalytic residues E184 and E189 are involved in retaining hydrolytic mechanisms. In-vitro, interaction with negatively charged carboxymethyl cellulose (CMC) enhanced the enzyme activity by 15% while positively charged chitosan (CS) decreased the enzyme activity by 12%, highlighting the effect of biopolymers on enzyme activity within the microenvironment interaction. These findings may provide insight into biochemical properties and application of WP_199597959 for agarose degradation.

Practical implementation of efficient biocatalysts for large-scale production of indigo remains challenging. Microbial cytochrome P450s may be useful for indigo production, but this has been rarely reported. We discovered that CYP105D18 catalysed H2O2-mediated C-3 hydroxylation of indole to synthesize indigo. A cell-free lysate from Escherichia coli containing CYP105D18 peroxygenase obtained after cell disruption was optimized for in vitro reaction. Next, 250 µM hydroxylamine was added to the cell-free lysate to inhibit other H2O2-utilizing enzymes that interfere with the CYP105D18 function. Furthermore, the active-site residues of CYP105D18, namely L87, A235, A282, and I386, involved in indole binding were mutated. L87F resulted in an approximately 12-fold increase in CYP105D18 activity. The catalytic efficiencies of the wild-type and L87F mutant were 0.01 and 0.12 mM-1min-1, respectively. Fed-batch fermentation using enriched autoinduction medium was used for higher production of E. coli cells containing CYP105D18 peroxygenase. The Cell-free lysate of disrupted cells yielded 710 mg/L of indigo in 20 min. This represents a simple enzymatic approach for indigo biosynthesis using cell-free lysate of E. coli overexpressing CYP105D18, H2O2, and catalase inhibitor without the need for multi enzyme systems and expensive cofactors. This single-enzyme system, used in a rapid process for indigo formation, could serve as an efficient approach for commercial bio-indigo production.

Gelidibacter salicanalis strains remain poorly understood in terms of their genomic attributes and are rarely reported for the degradation of polysaccharides within the niche. Gelidibacter salicanalis PAMC21136, an Antarctic isolate, was sequenced and functionally annotated. 251 genes were classified into the CAZyme families, which includes the highest number of GH family, highlighting peculiarity towards various macro biopolymers. Further, based on domain architecture and multiple sequence alignment, gene belonging to the family18 glycoside hydrolase was identified as chitinase. The gene encoding chitinase (MBJ7879808.1) was successfully cloned and expressed in Escherichia coli BL21 cells. MBJ7879808.1 demonstrated activity towards colloidal chitin but there was no detectable activity towards microcrystalline crab shell chitin. Enzyme exhibited an exo-pattern of hydrolysis on both colloidal chitin, and shorter chain GlcNAc oligomers namely [(GlcNAc)3, (GlcNAc)4, (GlcNAc)5, and (GlcNAc)6] producing GlcNAc as a single final product. Biochemical characterization revealed an optimum activity at 25 °C and pH 7.0. Mn²⁺ ions significantly enhanced the activity by 1.7-fold. Turnover number (Kcat) was 8.6 min−1 and catalytic efficiency (Kcat/Km) was 2.0ml/mg/min towards the substrate colloidal chitin. These findings expanded the polysaccharide degradation capability of Gelidibacter salicanalis PAMC21136 and highlight the biotechnological potential of the GH18 enzyme towards production of chitooligomers.

Cytochrome P450 monooxygenases perform a multitude of roles, including the generation of hydroxylated aromatic compounds that might be utilized by microorganisms for their survival. WGS data of Amycolatopsis magusensis KCCM40447 revealed a complete circular genome of 9,099,986 base pairs and functionally assigned 8601 protein-encoding genes. Genomic analysis confirmed that the gene for 4-methoxybenzoate monoxygenase (CYP199A35) was conserved in close proximity to the gene for 4-hydroxybenzoate transporter (PcaK). The co-localized genes encoding CYP199A35, and ferredoxin-NAD(P) reductase (Mbr) represent a two-component system for electron transfer. CYP199A35 was specific for O-demethylation of para O-methyl substituted benzoic acid derivatives, 4-methoxybenzoate (4 MB), and 4-methoxycinnamic acid (4MCA) using the native redox partner (Mbr); two-component system and non-physiological redox partners (Pdr/Pdx); three-component system. The catalytic efficiency for O-demethylation of 4 MB using Mbr and Pdr/Pdx was 0.02 ± 0.006 min−1 μM−1 and 0.07 ± 0.02 min−1 μM−1 respectively. Further, sequence annotation and function prediction by RAST and KEEG analysis revealed a complete catabolic pathway for the utilization of 4 MB by strain KCCM40447, which was also proved experimentally.

Heavy metals, including mercury, are non-biodegradable and highly toxic to microorganisms even at low concentrations. Understanding the mechanisms underlying the environmental adaptability of microorganisms with Hg resistance holds promise for their use in Hg bioremediation. We characterized Gbs MerA, a mercury reductase belonging to the mercury-resistant operon of Gelidibacter salicanalis PAMC21136, and found its maximum activity of 474.7 µmol/min/mg in reducing Hg +2 . In the presence of Ag and Mn, the enzyme exhibited moderate activity as 236.5 µmol/min/mg and 69 µmol/min/mg, respectively. Gbs MerA exhibited optimal activity at pH 7.0 and a temperature of 60 °C. Moreover, the crystal structure of Gbs MerA and structural comparison with homologues indicated that Gbs MerA contains residues, Tyr437´ and Asp47, which may be responsible for metal transfer at the si -face by providing a hydroxyl group (−OH) to abstract a proton from the thiol group of cysteine. The complex structure with NADPH indicated that Y174 in the re -face can change its side chain direction upon NADPH binding, indicating that Y174 may have a role as a gate for NADPH binding. Moreover, the heterologous host expressing Gbs MerA (pGbsMerA) is more resistant to Hg toxicity when compared to the host lacking Gbs MerA. Overall, this study provides a background for understanding the catalytic mechanism and Hg detoxification by Gbs MerA and suggests the application of genetically engineered E. coli strains for environmental Hg removal.

CYP105D18 supports H2O2 as an oxygen surrogate for catalysis well and shows high H2O2 resistance capacity. We report the hydroxylation of different steroids using H2O2 as a cosubstrate. Testosterone was regiospecifically hydroxylated to 2β-hydroxytestosterone. Based on the experimental data and molecular docking, we predicted that hydroxylation of methyl testosterone and nandrolone would occur at position 2 in the A-ring, while hydroxylation of androstenedione and adrenosterone was predicted to occur in the B-ring. Further, structure-guided rational design of the substrate access channel was performed with the mutagenesis of residues S63, R82, and F184. Among the mutants, S63A showed a marked decrease in product formation, while F184A showed a significant increase in product formation in testosterone, nandrolone, methyl testosterone, androstenedione, and adrenosterone. The catalytic efficiency (kcat/Km) toward testosterone was increased 1.36-fold in the F184A mutant over that in the wild-type enzyme. These findings might facilitate the potential use of CYP105D18 and further engineering to establish the basis of biotechnological applications. IMPORTANCE The structural modification of steroids is a challenging chemical reaction. Modifying the core ring and the side chain improves the biological activity of steroids. In particular, bacterial cytochrome P450s are used as promiscuous enzymes for the activation of nonreactive carbons of steroids. In the present work, we reported the H2O2-mediated hydroxylation of steroids by CYP105D18, which also overcomes the use of expensive cofactors. Further, exploring the substrate access channel and modifying the bulky amino acid F184A increase substrate conversion while modifying the substrate recognizing amino acid S63 markedly decreases product formation. Exploring the substrate access channel and the rational design of CYP105D18 can improve the substrate conversion, which facilitates the engineering of P450s for industrial application.

The genomic analysis of Streptomyces sp. KCCM12257 presented 233 CAZyme genes with a predominant glycosyl hydrolase family. This contributes degradation of various polysaccharides including chitin and chitosan, and other promising candidates for the production of different oligosaccharides. We screened the strain providing different polysaccharides as a sole source of carbon and strain KCCM12257, showed higher activity towards colloidal chitosan. Further, we identified and characterized a new chitosanase (MDI5907146) of GH46 family. There was no activity towards chitin, carboxymethylcellulose, or even with chitosan powder. This enzyme acts on colloidal chitosan and hydrolyzes it down into monoacetyl chitobiose, which consists of two glucosamine units with an acetyl group attached to them. The maximum enzyme activity was observed at pH 6.5 and 40 °C using colloidal chitosan as a substrate. The Co2+ metal ions almost double the reaction as compared to other metal ions. The dissociation constant (Km) and of colloidal chitosan (≥90 % and ≥75%DD) were 3.03 mg/ml and 5.01 mg/ml respectively, while maximum velocity (Vmax) values were found to be 36 mg/ml, and 30 μM/μg/min, respectively. Similarly, catalytic efficiency (Kcat/Km) of colloidal chitosan with ≥90 %DD was 1.9 fold higher than colloidal chitosan with ≥75%DD.

Limited numbers of CYPs have been reported to work naturally as peroxygenases. The peroxide shunt pathway can be efficiently used as an alternative for the NAD(P)H and reductase systems, particularly in high hydrogen peroxide (H2O2) resistance CYPs. We reported the structural and biochemical features of CYP105D18 peroxygenase for its high H2O2 tolerance capacity. Q348 was a crucial residue for the stability of CYP105D18 during the exposure to H2O2. In addition, the role of the hydrophilic amino acid T239 from the I helix for peroxygenation and regiospecificity toward testosterone was investigated. Interestingly, T239E differs in product formation from wild type, catalyzing testosterone to androstenedione in the presence of H2O2. The other variant, T239A, worked with the Pdx/Pdr system and was unable to catalyze testosterone conversion in the presence of H2O2, suggesting the transformation of peroxygenase into monooxygenase. CYP105D18 supported the alternative method of H2O2 used for the catalysis of testosterone. The use of the same concentration of urea hydrogen peroxide adducts in place of direct H2O2 was more efficient for 2β-hydroxytestosterone conversion. Furthermore, in situ H2O2 generation using GOx/glucose system enhanced the catalytic efficiency (kcat/Km) for wild type and F184A by 1.3- and 1.9-fold, respectively, compared to direct use of H2O2 The engineering of CYP105D18, its improved peroxygenase activity, and alteration in the product oxidation facilitate CYP105D18 as a potential candidate for biotechnological applications.

Background. Clinical laboratory diagnosis and prognosis for diabetes mellitus is performed using blood as a major specimen; however, saliva may represent as an alternative noninvasive specimen of choice. This study aims to evaluate salivary biochemical parameters in diabetic and healthy individuals to substantiate saliva's role in the diagnosis and prognosis of type 2 diabetes mellitus (T2DM). Methods. This case-control study included 150 T2DM patients and 150 apparently healthy individuals. Socio-demographic data and anthropometric measurements were recorded using a standard questionnaire. Correlation between salivary and blood levels for each parameter was determined using Pearson correlation. Linear regression was performed to estimate the blood levels of the parameters from their salivary levels. Receiver operating characteristics (ROC) analysis was done to determine the diagnostic ability of salivary glucose and establish a sensitivity, specificity, and cut-off value. Results. Salivary glucose, TC, LDL-C, urea, and creatinine were significantly higher in people with diabetes than in the control population (p < 0.05). A significant positive correlation was found between salivary and blood parameters including glucose, TC, TG, LDL-C, urea, and creatinine except for HDL-C in both case and control groups. The linear relationship for each parameter, except glucose in case population and HDL-C in case, control, and the total population was observed between blood and saliva. ROC analysis gave a cut-off value of 1.9mg/dl for salivary glucose with 71.4% sensitivity and 72.3% specificity. Conclusion. Salivary estimation significantly reflects the blood parameters in this study, indicating that saliva can be a noninvasive specimen for the diagnosis and prognosis of T2DM.

The chitobiose (chb) operon is involved in the synthesis of chitooligosaccharide and is comprised of a BCARFG gene cluster. ChbG encodes a chitooligosaccharide deacetylase (CDA) which catalyzes the removal of one acetyl group from N,N'-diacetylchitobiose. It is considered a novel type of CDA due to its lack of sequence homology. Although there are various structural studies of CDAs linked to the kinetic properties of the enzyme, the structural information of ChbG is unavailable. In this study, the crystal structure of ChbG from Klebsiella pneumoniae is provided. The molecular basis of deacetylation of diacetylchitobiose by ChbG is determined based on structural analysis, mutagenesis, biophysical analysis, and in silico docking of the substrate, diacetylchitobiose. This study contributes towards a deeper understanding of chitin and chitosan biology, as well as provides a platform to engineer CDA biocatalysts.

This study aims to assess vitamin D deficiency-induced dyslipidemia and cardiovascular disease (CVD) risk in poor glycemic control among type 2 diabetes mellitus (T2DM) patients. This study was carried out among 455 T2DM patients involving poor glycemic control ( n = 247) and good glycemic control ( n = 208). Fasting plasma glucose (FPG) and HbA 1 c were measured to assess glycemic control. Cardiac risk ratio, atherogenic index plasma, and atherogenic coefficient were calculated to assess and compare the CVD risk in different groups. Patients with poor control had a significantly higher level of total cholesterol (TC), triglyceride (TG), and non-high-density lipoprotein lipase cholesterol (non-HDL-C), atherogenic variables, and lower level of high-density lipoprotein lipase cholesterol (HDL-C) as compared to patients with good glycemic control. We also observed significant negative correlation of vitamin D with lipid markers and atherogenic variables in poor glycemic control diabetic population. The serum vitamin D levels were inversely associated with HbA 1 c, FPG, TG, TC, and non-HDL-C. Furthermore, hypercholesterolemia, hypertriglyceridemia, and elevated non-HDL-C were the independent risks in hypovitaminosis D population. Vitamin D deficiency in poor glycemic control is likely to develop dyslipidemia as compared to vitamin D insufficient and sufficient groups. Thus, vitamin D supplementation and an increase in exposure to sunlight may reduce the risk of cardiovascular complications in diabetes.

The bacterial CYP105 family is involved in secondary metabolite biosynthetic pathways and plays essential roles in the biotransformation of xenobiotics. This study investigates the newly identified H2O2-mediated CYP105D18 from Streptomyces laurentii as the first bacterial CYP for N-oxidation. The catalytic efficiency of CYP105D18 for papaverine N-oxidation was 1.43 s−1 µM−1. The heme oxidation rate (k) was low (<0.3 min−1) in the presence of 200 mM H2O2. This high H2O2 tolerance capacity of CYP105D18 led to higher turnover prior to heme oxidation. Additionally, the high-resolution papaverine complexed structure and substrate-free structure of CYP105D18 were determined. Structural analysis and activity assay results revealed that CYP105D18 had a strong substrate preference for papaverine because of its bendable structure. These findings establish a basis for biotechnological applications of CYP105D18 in the pharmaceutical and medicinal industries.

7α-Hydroxysteroid dehydrogenase (7α-HSDH) catalyzes the dehydrogenation of a hydroxyl group at the 7α position in steroid substrates using NAD+ or NADP+ as a co-factor. Although studies have determined the binary and ternary complex structures, detailed structural changes induced by ligand and co-factor binding remain unclear, because ligand-free structures are not yet available. Here, we present the crystal structure of apo 7α-HSDH from Escherichia coli (Eco-7α-HSDH) at 2.7 Å resolution. We found that the apo form undergoes substantial conformational changes in the β4-α4 loop, α7-α8 helices, and C-terminus loop among the four subunits comprising the tetramer. Furthermore, a comparison of the apo structure with the binary (NAD+)-complex and ternary (NADH and 7-oxoglycochenodeoxycholic acid)-complex Eco-7α-HSDH structures revealed that only the ternary-complex structure has a fully closed conformation, whereas the binary-complex and apo structures have a semi-closed or open conformation. This open-to-closed transition forces several catalytically important residues (S146, Y159, and K163) into correct positions for catalysis. To confirm the catalytic activity, we used alcohol dehydrogenase for NAD+ regeneration to allow efficient conversion of chenodeoxycholic acid to 7-ketolithocholic acid by Eco-7α-HSDH. These findings demonstrate that apo Eco-7α-HSDH exhibits intrinsically flexible characteristics with an open conformation. This structural information provides novel insight into the 7α-HSDH reaction mechanism.

Objective: Diabetes is a chronic metabolic disease which is growing at worrying rate in developing country like Nepal. . It is prevalent in children, adolescent and young adults due to increase in risk factors such as over feeding, physical inactivity, obesity. Therefore, the aim of this study was to assess diabetes risk score among young individuals. Methods: Across sectional study was conducted among students of age group (18to 25 years) studying at Manmohan Memorial Institute of Health Sciences and Central Institute of Science and Technology. All the socio-demographic data, anthropometric measurements, lifestyle and dietary habits were recorded by using standard questionnaire. Fasting plasma glucose and lipid profile were estimated by using standard manufactthe urer’s guideline. Then the Diabetes risk score was calculated by The Finnish Diabetes Risk Score (FINDRISC tool). Association between cardio-metabolic risk factors and diabetes risk score was established by ANOVA test. Results: Among total 825 students, 739 (89.6%) were recruited for the study with complete questionnaire, anthropometric measurement and fasting blood sample. Median age of the study population was 20 years. Among total study population, 553 (74.80%) were under low risk (FINDRISC<7), 164 (22.18%) were at slightly elevated risk (FINDRISC 7-11), 15 (2.02%) were at moderate risk (FINDRISC 12-14) and 7 (1.01%) were at high risk of diabetes. BMI, TC and LDL-C were higher at significant level (p<0.001) with increasing diabetes risk score in our study population. Conclusion: Risk factors for diabetes were common among young adults. Early assessment of diabetes risk in young may provide insights for preventive and control plan for risk population.

Aims: This study aimed to find the association of liver biomarker with diabetes population. Also, this study focused to find out the risk factors associated with liver disease in Nepalese diabetes patient. Methods: This study was carried out among 238 diabetes and 238 apparently healthy population who attended Modern diagnostic laboratory and Research center, Kathmandu, Nepal. HbA1c and fasting plasma glucose was measure to assess the diabetes population and glycemic control. Aspartate to platelet ratio index (APRI), gamma-glutamyl transpeptidase to platelet ratio (GPR), fibrosis-4 index (FIB-4), and triglyceride and glucose index (TyG) were assess for prediction of hidden risk liver disease. Diabetic patient with higher than the cut-off value obtained from ROC curve analysis of different liver marker index were subjected to multivariate regression analysis to measure the independent risk factor for progression liver disease in different model. Results: Patient with poor glycemic control had a significantly higher level of APRI (P=0.05), GPR (P=0.039), and TyG (P<0.001). Higher HbA1c showed significantly positive correlation with APRI (r=0.154, P=0.017), GPR (r=0.203, P=0.002), FIB4 (r=0.132, P=0.042), and TyG (r=0.510, P<0.001) in diabetic population. The Area under ROC curve of GPR was 0.700 (0.654-0.747), APRI 0.839 (0.803-0.874), FIB-4 0.820 (0.783-0.857), and TyG 0.909 (0.882-0.874) with p-value <0.05. The cut-off value (sensitivity, specificity) of GPR was 0.227 (63.4%, 63%), APRI 0.241 (71.0%, 80.3%), FIB-4 1.65 (71.8%, 77.3%), and TyG 8.85 (79%, 93.3%) respectively. Triglyceride, AST, and GGT was independent risk factor followed same trend in different 4 model while HbA1c and ALT showed independent risk factor in 3 models. Conclusions: APRI, GPR, FIB4 and TyG can define the hidden risk liver disease in T2DM. The independent risk factors for progression of liver disease in those population are hypertriglyceridemia, higher AST, and higher GGT. Routinely screening for markers may prevent progression of liver disease in T2DM patients.

Background: The widespread dissemination of unhealthy dietary habits, childhood-teenage obesity, and sedentary lifestyle in young adults has paved the way for public health burden metabolic syndrome and early onset of type 2 diabetes mellitus. The aim of this study was to assess the prevalence and risk factors for metabolic syndrome and diabetes among young adult students. Methods: This cross-sectional study was conducted among students of age group (18 to 25 years) studying at Manmohan Memorial Institute of Health Sciences and Central Institute of Science and Technology. The diabetes risk score of each individual was calculated by the Finnish Diabetes Risk Score (FINDRISC tool). Independent risk factors for diabetes and metabolic syndrome were measured by multivariable logistic regression analysis. The p-value of <0.05 was considered statistically significant in this study. Results: A total of 825 students were recruited and 739 (89.6%) students completed the study with all the fulfilled criteria. The metabolic syndrome (Harmonized Joint Scientific Statement (HJSS) criteria) was present in 7.1%, and the most prevalent defining component was low HDL-C (78%); 74.8% of students were under low risk, 22.18% were at slightly elevated risk, 2.02% were at moderate risk, and 1.01% were at high risk of diabetes. The cardiometabolic risk factors like BMI, TC, and LDL-C were higher at a significant level (p<0.001) with an increased diabetes risk score. Independent lifestyle risk factor for metabolic syndrome was current smoking (AOR, 4.49, 95% CI 1.38-14.62) whereas, an independent lifestyle risk factor for diabetes was low adherence to physical exercise (AOR, 4.81, 95% CI, 2.90-7.99). Conclusion: Metabolic syndrome is present, although in low numbers in young adults putting them at risk to develop diabetes in the near future. Early assessment of metabolic syndrome and diabetes risk in young may provide insights for preventive and control plans for risk population.

Sushant Pokhrel 1,2 Bashu Dev Pardhe 1,3 Nisha Giri Rakesh Pokhrel Deliya Paudel 1Department of Laboratory Medicine, Manmohan Memorial Institute of Health Sciences, Kathmandu, Nepal; 2Modern Diagnostic Laboratory and Research Center, Kathmandu, Nepal; 3Department of Life Science and Biochemical Engineering, Sun Moon University, AsanSi, Chungnam, South Korea; 4Institute of Medicine, Tribhuvan University, Kathmandu, Nepal; 5Manmohan Memorial Medical College and Teaching Hospital, Kathmandu, Nepal Abstract: Dermatomyositis is an idiopathic myopathy involving progressive muscle weakness with skin manifestation. Diagnosis is based upon the progressive muscle weakness, skin rashes, elevated serum muscle enzymes, muscle biopsy, and abnormal electromyogram. The incidence of dermatomyositis is rare. In this case study, we report a case of classical dermatomyositis without incidence of malignancy. Timely diagnosis and administration of steroid led to better prognosis of the patient.

Purpose The patient believes in adherence to medication rather than to self-care adherence and lifestyle changes for the management of diabetes. This study was carried out to establish the association of self-care adherence and their barriers in poor glycemic control in our diabetic population. Patients and methods This cross-sectional study was conducted among 480 already diagnosed diabetes outpatients attended in our two hospitals. Glycaemic control was defined by levels of HbA1c. Socio-demographic data, lifestyle variables and anthropometric measurements were recorded using a standard questionnaire. Fasting blood glucose, HbA1c and lipid profiles were estimated using the manufacturer’s guideline. Student’s t-test and one-way ANOVA were used for comparison between different groups and the correlation was established by Spearman correlation. Risk factors associated with poor glycaemic control were verified by logistic regression analysis. Results The mean HbA1c of the study population was 7.4±1.3% and 65.4% had poor glycaemic control with mean 8.0±1.1%. Higher HbA1c levels were significantly associated with duration of diabetes, a number of drugs used, patient–physician relationship and knowledge about diabetes. The poor glycaemic control was significantly associated with low adherence of following the meal plan, regular medication and regular exercising (p<0.001). Among all the barriers, a too busy schedule for following the meal plan, taking medications and exercising regularly was significantly correlated with HbA1c levels. Multivariable logistic regression analysis showed irregular meal plan (OR=5.27), irregular exercise (OR=2.25), number of medication used (OR= 0.19) and lesser extent patient–physician relationship (OR=2.68) were independent risk factors for poor glycaemic control. Conclusion The poor glycaemic control was associated with poor adherence to self-care adherence and their barriers in our diabetic population. Integrated knowledge on diabetes management should be targeted to improve glycaemic control in our communities.