Naringin is a main flavonoid in citrus industry,and it is also a bitter matter in fruit juice.In our previous work,a mutant strain can produceα-L-rhamnosidase with good hydrolysis efficiency for hesperidin other than...Naringin is a main flavonoid in citrus industry,and it is also a bitter matter in fruit juice.In our previous work,a mutant strain can produceα-L-rhamnosidase with good hydrolysis efficiency for hesperidin other than naringin.To extend its function based on our present study,we have explored the reason why the efficiency of catalysing hesperidin into hesperetin 7-O-glucoside than catalysing naringin into prunin.As a result,we found five key amino acid residues were responsible for the difference by molecular docking and molecular dynamics simulations.The terminal rhamnose of naringin is bound to the inside of the active pocket of the enzyme,and the addition of two new hydrogen bonds come out after the hydrolysis of naringin.So the active pocket began to contract,making it difficult for the terminal rhamnose to detach from it.5 key amino acid residues,THR203,GLU158,LYS276,GLN133,and ILE258 were identified,and obtained five new mutants by mutating them into ALA.Among them,the mutant strain of K276A showed higher conversion efficiency from naringin to prunin along with higher thermal stability.This work not only improved the affinity ofα-L-Rhamnosidase for catalysing bittering matter of naringin and enhances its potential application in the food and pharmaceutical industries,but also provided an effective technological strategy for improving the affinity of other enzymes.展开更多
基金support of the Science and Technology Planning Project of Agricultural Division 9 in Xinjiang Production and Construction Corps.,P.R.China(2023JS020)the Science and Technology Planning Project in Guangdong,P.R.China(SDZX2022041,SDZX2023036 and SDZX2023033)+2 种基金the Science and Technology Innovation Strategy Special Project in Guangdong Province,P.R.China(pdjh 2023a0025)the Key R&D Program Projects in Guangdong,P.R.China(2023B0202040002 and 2024B0202010001)the‘Open Competition’Science&Technology Research Project of National Excellent Engineer Innovation Research Institute for Advanced Manufacturing Industry in Guangdong Hong Kong Macao Greater Bay Area(Foshan)(JBGS2024003).
文摘Naringin is a main flavonoid in citrus industry,and it is also a bitter matter in fruit juice.In our previous work,a mutant strain can produceα-L-rhamnosidase with good hydrolysis efficiency for hesperidin other than naringin.To extend its function based on our present study,we have explored the reason why the efficiency of catalysing hesperidin into hesperetin 7-O-glucoside than catalysing naringin into prunin.As a result,we found five key amino acid residues were responsible for the difference by molecular docking and molecular dynamics simulations.The terminal rhamnose of naringin is bound to the inside of the active pocket of the enzyme,and the addition of two new hydrogen bonds come out after the hydrolysis of naringin.So the active pocket began to contract,making it difficult for the terminal rhamnose to detach from it.5 key amino acid residues,THR203,GLU158,LYS276,GLN133,and ILE258 were identified,and obtained five new mutants by mutating them into ALA.Among them,the mutant strain of K276A showed higher conversion efficiency from naringin to prunin along with higher thermal stability.This work not only improved the affinity ofα-L-Rhamnosidase for catalysing bittering matter of naringin and enhances its potential application in the food and pharmaceutical industries,but also provided an effective technological strategy for improving the affinity of other enzymes.
