title: Cloning and purification of the first termicin-like peptide from the cockroach Eupolyphaga sinensis.
abstract: BACKGROUND
  Termicin is an antimicrobial peptide with six cysteines forming three disulfide bridges that was firstly isolated from the salivary glands and hemocytes of the termite Pseudacanthotermes spiniger. In contrast to many broad-spectrum antimicrobial peptides, termicin is most active against filamentous fungi. Although more than one hundred complementary DNAs (cDNAs) encoding termicin-like peptides have been reported to date, all these termicin-like peptides were obtained from Isoptera insects.
  
  
  METHODS
  The cDNA was cloned by combination of cDNA library construction kit and DNA sequencing. The polypeptide was purified by gel filtration and reversed-phase high performance liquid chromatography (RP-HPLC). Its amino acid sequence was determined by Edman degradation and mass spectrometry. Antimicrobial activity was tested against several bacterial and fungal strains. The minimum inhibitory concentration (MIC) was determined by microdilution tests.
  
  
  RESULTS
  A novel termicin-like peptide with primary structure ACDFQQCWVTCQRQYSINFISARCNGDSCVCTFRT was purified from extracts of the cockroach Eupolyphaga sinensis (Insecta: Blattodea). The cDNA encoding Es-termicin was cloned by cDNA library screening. This cDNA encoded a 60 amino acid precursor which included a 25 amino acid signal peptide. Amino acid sequence deduced from the cDNA matched well with the result of protein Edman degradation. Susceptibility test indicated that Es-termicin showed strong ability to kill fungi with a MIC of 25 μg/mL against Candida albicans ATCC 90028. It only showed limited potency to affect the growth of Gram-positive bacteria with a MIC of 200 μg/mL against Enterococcus faecalis ATCC 29212. It was inactive against gram-negative bacteria at the highest concentration tested (400 μg/mL). Es-termicin showed high sequence similarity with termicins from many species of termites (Insecta: Isoptera).
  
  
  CONCLUSIONS
  This is the first report of a termicin-like peptide isolated from E. sinensis that belongs to the insect order Blattodea. Our results demonstrate the diversity of termicin-like peptides, as well as antimicrobial peptides in insects.
authors.affiliation: Key Laboratory of Special Biological Resource Development and Utilization of Universities in Yunnan Province, Key Lab of Aquatic Ecological Restoration of Dianchi Lake in Kunming, Department of Biological Science and Technology, Kunming University, Kunming, 650214 China.
authors.forename: Zichao
authors.identifier: 0000-0002-7509-6209
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Oncology, Yan'an Hospital of Kunming City; Yunnan, Cardiovascular Hospital; and Yan'an Hospital of Kunming Medical University, Kunming, 650051 China.
authors.forename: Kehua
authors.initials: K
authors.lastname: Yuan
authors.affiliation: Department of Obstetrics and Gynecology, Shenzhen Maternal and Child Health Care Hospital, Affiliated to Southern Medical University, Shenzhen, 518028 China.
authors.forename: Ruopeng
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Special Biological Resource Development and Utilization of Universities in Yunnan Province, Key Lab of Aquatic Ecological Restoration of Dianchi Lake in Kunming, Department of Biological Science and Technology, Kunming University, Kunming, 650214 China.
authors.forename: Xuchen
authors.initials: X
authors.lastname: Ren
authors.affiliation: Key Laboratory of Special Biological Resource Development and Utilization of Universities in Yunnan Province, Key Lab of Aquatic Ecological Restoration of Dianchi Lake in Kunming, Department of Biological Science and Technology, Kunming University, Kunming, 650214 China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Liu
authors.affiliation: Yunnan Key Laboratory of Fertility Regulation and Minority Eugenics, Yunnan Population and Family Planning Research Institute, Kunming, 650021 China.|First Affiliated Hospital of Kunming Medical University, Xichang Road 295#, Kunming, Yunnan 650032 China.
authors.forename: Shuhua
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Special Biological Resource Development and Utilization of Universities in Yunnan Province, Key Lab of Aquatic Ecological Restoration of Dianchi Lake in Kunming, Department of Biological Science and Technology, Kunming University, Kunming, 650214 China.|Kunming University, Puxin Road 2#, Kunming, Yunnan 650214 China.
authors.forename: Dingkang
authors.initials: D
authors.lastname: Wang
keywords: Antifungal peptide; Cockroach; Es-termicin; Eupolyphaga sinensis; Termicin-like peptide
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