Characterization of D-lyxose isomerase from different sources
學生姓名:
邱琬軒
指導教授:
方翠筠
學期:
112上
摘 要:
D-lyxose isomerase is an aldose-ketose isomerase with broad substrate specificity that can catalyze isomerization reaction between enzymes, and has been used for the production of functional rare sugars D-mannose and L-ribose. At present, broad application of D-mannose
has been used in the field of food, cosmetic and pharmaceutical industries. The preparation of D-mannose derived from the chemical synthesis, plant extraction and enzymatic method, with the enzymatic approach being suited for industrial production. Therefore, this study aims to investigate characterization analysis of D-lyxose isomerase from different sources. A novel D19 lyxose isomerase (D-LIase) from Thermoprotei archaeon through genetic transformation and purification by using a nickel ion-affinity column has been used for analysis of optimal
conditions and substrate specificity. The results showed that the optimal conditions for D-LIase were at pH 6.5 and temperature 80 to 85°C, in the presence of 0.5 mM Ni2+ 22 . Furthermore, the conversion rate still reached approximately 20% when the reaction happened at 80 °C. This demonstrates the potential of D-LIase exhibits excellent substrate specificity at high temperatures. On the other hand, D-lyxose isomerase from Bacillus velezensis(BvLI) exhibited maximum activity at 55°C and pH 6.5. The addition of Co2+ and Mn2+ 26 effectively enhanced BvLI activity, while Cu2+ and Zn2+ 27 completely inhibited its activity. The optimal conditions for producing D-mannose were achieved by adding 500 g/L of D-fructose and 25 U/mL of the recombinant BvLI. According to these papers, both of the recombinant D-lyxose isomerases indicated highly thermal stability and suitable for survival in weakly acidic condition. In summary, D-lyxose isomerase have great potential for large-scale application in the production of rare sugars in the future.
has been used in the field of food, cosmetic and pharmaceutical industries. The preparation of D-mannose derived from the chemical synthesis, plant extraction and enzymatic method, with the enzymatic approach being suited for industrial production. Therefore, this study aims to investigate characterization analysis of D-lyxose isomerase from different sources. A novel D19 lyxose isomerase (D-LIase) from Thermoprotei archaeon through genetic transformation and purification by using a nickel ion-affinity column has been used for analysis of optimal
conditions and substrate specificity. The results showed that the optimal conditions for D-LIase were at pH 6.5 and temperature 80 to 85°C, in the presence of 0.5 mM Ni2+ 22 . Furthermore, the conversion rate still reached approximately 20% when the reaction happened at 80 °C. This demonstrates the potential of D-LIase exhibits excellent substrate specificity at high temperatures. On the other hand, D-lyxose isomerase from Bacillus velezensis(BvLI) exhibited maximum activity at 55°C and pH 6.5. The addition of Co2+ and Mn2+ 26 effectively enhanced BvLI activity, while Cu2+ and Zn2+ 27 completely inhibited its activity. The optimal conditions for producing D-mannose were achieved by adding 500 g/L of D-fructose and 25 U/mL of the recombinant BvLI. According to these papers, both of the recombinant D-lyxose isomerases indicated highly thermal stability and suitable for survival in weakly acidic condition. In summary, D-lyxose isomerase have great potential for large-scale application in the production of rare sugars in the future.
邱琬軒.pdf