Production of small antibacterial peptides using silkworm-baculovirus protein expression system

Mai Fukushima, Kazuhiro Iiyama, Jun Yamashita, Masutaka Furue, Gaku Tsuji, Shigeo Imanishi, Hiroaki Mon, Man Lee, Takahiro Kusakabe

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The recombinant proteins with strong antimicrobial activity are known to be very difficult to express using bacterial expression system. Here, human β-defensin (DEFB) 1, DEFB2, and DEFB3 were successfully produced using a silkworm-baculovirus protein expression system. We have generated four baculoviruses for each DEFB protein to compare the effect of different peptide tags in secretion into silkworm larval hemolymph. Interestingly, the best performing peptide tags for the secretion were different among DEFBs: C-terminal GST-H8 tag for DEFB1, N-terminal H8 tag for DEFB2, and C-terminal H8 tag for DEFB3, respectively. In addition, the colony count assay demonstrated that the recombinant DEFB2 s showed antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, and Paenibacillus thiaminolyticus.

Original languageEnglish
Pages (from-to)565-576
Number of pages12
JournalPreparative Biochemistry and Biotechnology
Volume43
Issue number6
DOIs
Publication statusPublished - Aug 18 2013

Fingerprint

Bombyx
Baculoviridae
Paenibacillus
Defensins
Peptides
Hemolymph
Recombinant Proteins
Pseudomonas aeruginosa
Escherichia coli
Assays
Proteins

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry

Cite this

Production of small antibacterial peptides using silkworm-baculovirus protein expression system. / Fukushima, Mai; Iiyama, Kazuhiro; Yamashita, Jun; Furue, Masutaka; Tsuji, Gaku; Imanishi, Shigeo; Mon, Hiroaki; Lee, Man; Kusakabe, Takahiro.

In: Preparative Biochemistry and Biotechnology, Vol. 43, No. 6, 18.08.2013, p. 565-576.

Research output: Contribution to journalArticle

@article{d6efa990207d482e8749b0c6ca590fb5,
title = "Production of small antibacterial peptides using silkworm-baculovirus protein expression system",
abstract = "The recombinant proteins with strong antimicrobial activity are known to be very difficult to express using bacterial expression system. Here, human β-defensin (DEFB) 1, DEFB2, and DEFB3 were successfully produced using a silkworm-baculovirus protein expression system. We have generated four baculoviruses for each DEFB protein to compare the effect of different peptide tags in secretion into silkworm larval hemolymph. Interestingly, the best performing peptide tags for the secretion were different among DEFBs: C-terminal GST-H8 tag for DEFB1, N-terminal H8 tag for DEFB2, and C-terminal H8 tag for DEFB3, respectively. In addition, the colony count assay demonstrated that the recombinant DEFB2 s showed antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, and Paenibacillus thiaminolyticus.",
author = "Mai Fukushima and Kazuhiro Iiyama and Jun Yamashita and Masutaka Furue and Gaku Tsuji and Shigeo Imanishi and Hiroaki Mon and Man Lee and Takahiro Kusakabe",
year = "2013",
month = "8",
day = "18",
doi = "10.1080/10826068.2012.762717",
language = "English",
volume = "43",
pages = "565--576",
journal = "Preparative Biochemistry and Biotechnology",
issn = "1082-6068",
publisher = "Taylor and Francis Ltd.",
number = "6",

}

TY - JOUR

T1 - Production of small antibacterial peptides using silkworm-baculovirus protein expression system

AU - Fukushima, Mai

AU - Iiyama, Kazuhiro

AU - Yamashita, Jun

AU - Furue, Masutaka

AU - Tsuji, Gaku

AU - Imanishi, Shigeo

AU - Mon, Hiroaki

AU - Lee, Man

AU - Kusakabe, Takahiro

PY - 2013/8/18

Y1 - 2013/8/18

N2 - The recombinant proteins with strong antimicrobial activity are known to be very difficult to express using bacterial expression system. Here, human β-defensin (DEFB) 1, DEFB2, and DEFB3 were successfully produced using a silkworm-baculovirus protein expression system. We have generated four baculoviruses for each DEFB protein to compare the effect of different peptide tags in secretion into silkworm larval hemolymph. Interestingly, the best performing peptide tags for the secretion were different among DEFBs: C-terminal GST-H8 tag for DEFB1, N-terminal H8 tag for DEFB2, and C-terminal H8 tag for DEFB3, respectively. In addition, the colony count assay demonstrated that the recombinant DEFB2 s showed antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, and Paenibacillus thiaminolyticus.

AB - The recombinant proteins with strong antimicrobial activity are known to be very difficult to express using bacterial expression system. Here, human β-defensin (DEFB) 1, DEFB2, and DEFB3 were successfully produced using a silkworm-baculovirus protein expression system. We have generated four baculoviruses for each DEFB protein to compare the effect of different peptide tags in secretion into silkworm larval hemolymph. Interestingly, the best performing peptide tags for the secretion were different among DEFBs: C-terminal GST-H8 tag for DEFB1, N-terminal H8 tag for DEFB2, and C-terminal H8 tag for DEFB3, respectively. In addition, the colony count assay demonstrated that the recombinant DEFB2 s showed antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, and Paenibacillus thiaminolyticus.

UR - http://www.scopus.com/inward/record.url?scp=84879101133&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84879101133&partnerID=8YFLogxK

U2 - 10.1080/10826068.2012.762717

DO - 10.1080/10826068.2012.762717

M3 - Article

C2 - 23742088

AN - SCOPUS:84879101133

VL - 43

SP - 565

EP - 576

JO - Preparative Biochemistry and Biotechnology

JF - Preparative Biochemistry and Biotechnology

SN - 1082-6068

IS - 6

ER -