Cell lysate of strain BL21 showed significant D-glucose isomerase activity. The rate of glucose conversion was increased up to 40% when cells were induced with 1% D-xylose. BL21 xylose isomerase (ECXI-BL21) was purified to homogeneity, up to 1.9-fold with overall 10.88% enzyme yield by heat shock, salting out and electro-elution. The molecular mass of ECXI-BL21 was estimated as 43.9 kDa on SDS-PAGE. pH and T of the enzyme were calculated as 7.0 and 50 °C, respectively. Activation energy ( ) of ECXI-BL21 was 45 kJ/mol. Enzyme was stable from 30 to 55 °C and at pH range 6.0-8.0. ECXI-BL21 was activated by 10 mM magnesium (35%), 0.5 mM cobalt (20%) and manganese (25%), and 0.5/10 mM Mn/Mg (50%) and Co/Mg (30%) as compared to ECXI-BL21. Catalytic affinity ( ) of ECXI-BL21 for D-glucose was calculated as 0.82 mM, while maximum velocity ( ) of the reaction D-glucose ⇌ D-fructose was 108 μmol/mg/min. D-fructose formed was identified on silica gel plate. This thermophilic enzyme, = 75 °C, has great potential for high fructose syrup production used in food and soft drink industries.
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| http://dx.doi.org/10.1007/s13205-019-2036-6 | DOI Listing |
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