Hyperglycemic activity of the recombinant crustacean hyperglycemic hormone B1 isoform (CHH-B1) of the Pacific white shrimp Litopenaeus vannamei
•rCHH-B1 from L. vannamei was cloned and expressed in Pichia.•rCHH-B1 was successfully purified by RP-HPLC as a bioactive peptide.•The identity of CHH-B1 was confirmed by immunodetection and N-terminal sequencing.•rCHH-B1 is a CHH-like peptide that showed high hyperglycemic activity.•rCHH-B1 display...
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Veröffentlicht in: | Peptides (New York, N.Y. : 1980) N.Y. : 1980), 2015-09, Vol.71, p.32-39 |
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Zusammenfassung: | •rCHH-B1 from L. vannamei was cloned and expressed in Pichia.•rCHH-B1 was successfully purified by RP-HPLC as a bioactive peptide.•The identity of CHH-B1 was confirmed by immunodetection and N-terminal sequencing.•rCHH-B1 is a CHH-like peptide that showed high hyperglycemic activity.•rCHH-B1 displayed dose-dependent hyperglycemic activity in ablated shrimp.
Crustacean hyperglycemic hormone (CHH) is the most abundant neuropeptide produced by the X-organ/sinus gland (XO/SG) complex in the crustacean eyestalk. CHH plays a principal role in the control of glucose metabolism. The CHH-B1 isoform is produced in the eyestalk of Litopenaeus vannamei by alternative splicing of the chhB gene and its cDNA sequence has revealed that this isoform has a non-amidated C-terminal residue (CHH-like peptide). In this work, a recombinant CHH-B1 (rCHH-B1) with a sequence identical to the native hormone was expressed in the methylotrophic yeast Pichia pastoris X-33 and purified from the culture medium by RP-HPLC. The identity of the purified rCHH-B1 was confirmed by N-terminal sequencing and by using an anti-CHH-B1 polyclonal antibody. An in vivo assay showed that the hyperglycemic effect was dependant of the dosage of rCHH-B1, and the maximal hyperglycemic response was obtained with 250pmol treatment. These results suggest that the amino acid sequence of the C-terminus and its correct structure are both important for the hyperglycemic activity of naturally occurring non-amidated CHH peptides, such as CHH-B1. CHH-B1 appears to be the first reported CHH-like peptide with significant hyperglycemic activity produced in the sinus gland of a penaeid shrimp. |
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ISSN: | 0196-9781 1873-5169 |
DOI: | 10.1016/j.peptides.2015.05.014 |