1. A Novel Pseudomonas geniculata AGE Family Epimerase/Isomerase and Its Application in D-Mannose Synthesis
- Author
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Ying Li, Sheng Chen, Zhan-Zhi Liu, and Jing Wu
- Subjects
0106 biological sciences ,Identification ,Health (social science) ,Subfamily ,d-mannose ,Plant Science ,Isomerase ,lcsh:Chemical technology ,01 natural sciences ,Health Professions (miscellaneous) ,Microbiology ,Chemical synthesis ,Article ,law.invention ,d-mannose isomerase ,Active center ,03 medical and health sciences ,law ,010608 biotechnology ,lcsh:TP1-1185 ,Enzyme kinetics ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,AGE family epimerase/isomerase ,Chemistry ,Substrate (chemistry) ,d<%2Fspan>-mannose+isomerase%22">d-mannose isomerase ,d<%2Fspan>-mannose%22">d-mannose ,Enzyme ,Biochemistry ,Recombinant DNA ,Food Science - Abstract
d-mannose has exhibited excellent physiological properties in the food, pharmaceutical, and feed industries. Therefore, emerging attention has been applied to enzymatic production of d-mannose due to its advantage over chemical synthesis. The gene age of N-acetyl-d-glucosamine 2-epimerase family epimerase/isomerase (AGEase) derived from Pseudomonas geniculata was amplified, and the recombinant P. geniculata AGEase was characterized. The optimal temperature and pH of P. geniculata AGEase were 60 °, C and 7.5, respectively. The Km, kcat, and kcat/Km of P. geniculata AGEase for d-mannose were 49.2 ±, 8.5 mM, 476.3 ±, 4.0 s&minus, 1, and 9.7 ±, 0.5 s&minus, 1·, mM&minus, 1, respectively. The recombinant P. geniculata AGEase was classified into the YihS enzyme subfamily in the AGE enzyme family by analyzing its substrate specificity and active center of the three-dimensional (3D) structure. Further studies on the kinetics of different substrates showed that the P. geniculata AGEase belongs to the d-mannose isomerase of the YihS enzyme. The P. geniculata AGEase catalyzed the synthesis of d-mannose with d-fructose as a substrate, and the conversion rate was as high as 39.3% with the d-mannose yield of 78.6 g·, L&minus, 1 under optimal reaction conditions of 200 g·, 1d-fructose and 2.5 U·, mL&minus, 1P. geniculata AGEase. This novel P. geniculata AGEase has potential applications in the industrial production of d-mannose.
- Published
- 2020
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