Purification and characterization of a novel intracellular α-amylase with a wide variety of substrates hydrolysis and transglycosylation activity from Paenibacillus sp. SSG-1.
Intracellular α-amylase was a special glycoside hydrolase in the cytoplasm. We cloned and expressed an intracellular α-amylase, Amy, from Paenibacillus sp. SSG-1. The recombinant enzyme was purified by metal-affinity chromatography, exhibited a molecular mass of 71.7 kDa. Amy exhibited unexpectedly sequence similarity and evolutionary relationships with alpha-glucanotransferase. The docked results of Amy with maltose showed it had similar catalytic residues with α-amylase and glucanotransferase. The substrate specificity experiment showed that Amy could hydrolyze typical substrates into glucose and maltose. It was noteworthy that Amy showed the catalytic capacity of cyclomaltodextrinase and pullulanase. Meanwhile, Amy could transfer sugar molecules and form maltotetraose upon the hydrolysis of substrates. These results indicated that Amy was a novel intracellular α-amylase with distinct catalytic ability characteristics of hydrolyzing glycogen/cyclodextrin/pullulan and transglycosylation. We deduced that Amy may play an important role in utilizing maltooligosaccharides that released from extracellular α-glucan or storage α-glucan (glycogen) in Paenibacillus sp. SSG-1.
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Qingrui
Q
Xu
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China; National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Yu
Y
Cao
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Xi
X
Li
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Lin
L
Liu
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Shishang
S
Qin
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Yuhao
Y
Wang
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Yi
Y
Cao
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China.
Hui
H
Xu
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China. Electronic address: qiaodairong@scu.edu.cn.
Dairong
D
Qiao
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D003505:Cyclodextrins / Q000378:metabolism*; D005936:Glucans / Q000378:metabolism*; D006004:Glycogen Debranching Enzyme System / Q000302:isolation & purification / Q000378:metabolism; D006867:Hydrolases / Q000302:isolation & purification / Q000378:metabolism; D006868:Hydrolysis; D056507:Paenibacillus / Q000201:enzymology*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D000516:alpha-Amylases / Q000302:isolation & purification* / Q000378:metabolism*
D001426:Bacterial Proteins; D003505:Cyclodextrins; D005936:Glucans; D006004:Glycogen Debranching Enzyme System; D011994:Recombinant Proteins; C009109:pullulan; C020569:4 alpha-glucanotransferase; D006867:Hydrolases; D000516:alpha-Amylases
Cyclomaltodextrinase; GH13 family; Glycogen; Intracellular α-amylase; Pullulanase; Transglycosylation