Cloning, expression, and characteristic analysis of the novel β‐galactosidase from silkworm, Bombyx mori
Summary β‐Galactosidase is a critical exoglycosidase involved in the hydrolysis of lactose, the modification and degradation of glycoprotein in vivo. In this study, the β‐galactosidase gene of silkworm (BmGal), whose cDNA comprises 11 exons and contains an intact ORF of 1,821 bp, was cloned. The pro...
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Published in | Genesis (New York, N.Y. : 2000) Vol. 59; no. 9; pp. e23446 - n/a |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Hoboken, USA
John Wiley & Sons, Inc
01.09.2021
Wiley Subscription Services, Inc |
Subjects | |
Online Access | Get full text |
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Summary: | Summary
β‐Galactosidase is a critical exoglycosidase involved in the hydrolysis of lactose, the modification and degradation of glycoprotein in vivo. In this study, the β‐galactosidase gene of silkworm (BmGal), whose cDNA comprises 11 exons and contains an intact ORF of 1,821 bp, was cloned. The protein sequence of BmGal showed high similarity with other known insect β‐galactosidases. No activity of the BmGal expressed in Escherichia coli or Pichia pastoris was detected while it was successfully expressed with high enzyme activity in baculovirus expression system in silkworm, and the electrophoresis result revealed that the BmGal showed activity in oligomer mode. Enzyme activity assay showed that its optimum pH was 8.4 and its optimum temperature was 40 °C. What is more, we found that iron ions can stimulate the activity of the enzyme while cobalt, nickel, or lead ions can inhibit its activity significantly. Besides, the temporal–spatial transcription pattern of the BmGal mRNA level was analyzed, which showed that BmGal was transcribed at the highest level in the fifth larval instar but relatively low level in the pupal and adult stage, and the highest transcriptional level of BmGal was found in testis among all the tissues concerned. |
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Bibliography: | Funding information Yongzhu Yi and Jialei Li should be considered joint first authors. Central Public‐interest Scientific Institution Basal Research Fund for Chinese Academy of Tropical Agricultural Sciences, Grant/Award Number: 1610392021012; National Natural Science Foundation of China, Grant/Award Numbers: 32002236, 32072796; State Key Laboratory of Integrated Pest Management, Grant/Award Number: ChineseIPM1504; The Agricultural Science and Technology Innovation Program (ASTIP) ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1526-954X 1526-968X |
DOI: | 10.1002/dvg.23446 |