Cloning and Expression of β-(1,3-1,4) Glucanase (Lichenase) Gene in <i>Bacillus subtilis</i> RSKK246 to create new Probiotic in aquaculture

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Çam Ö., Baylan M., Mazi G.

ANAIS DA ACADEMIA BRASILEIRA DE CIENCIAS, cilt.94, sa.4, ss.1-8, 2022 (SCI-Expanded, Scopus)

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 94 Sayı: 4
  • Basım Tarihi: 2022
  • Doi Numarası: 10.1590/0001-3765202220200913
  • Dergi Adı: ANAIS DA ACADEMIA BRASILEIRA DE CIENCIAS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Aerospace Database, Animal Behavior Abstracts, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, CAB Abstracts, Communication Abstracts, EMBASE, INSPEC, MEDLINE, Metadex, Veterinary Science Database, zbMATH, Directory of Open Access Journals, Civil Engineering Abstracts
  • Sayfa Sayıları: ss.1-8
  • Anahtar Kelimeler: Bacillus subtilis, ss (1,3-1,4) glucanase, gene transfer, probiotic
  • Açık Arşiv Koleksiyonu: AVESİS Açık Erişim Koleksiyonu
  • Çukurova Üniversitesi Adresli: Evet

Özet

 β-Glucan is an essential component of the cell walls of grains such as oats

and barley. 1,3-1,4-β-D-glucan 4-glucanhydrolase (β-glucanase or lichenase) (EC 3.2.1.73)

is an enzyme with the ability to hydrolyze β-glucans. In this research, β-Glucan which

is a good source of feed additive fi sh probiotics, was used in order to benefi t from feed

quality in fi shery products, to increase live weight gain and to strengthen the immune

system. In this study, recombinant vector pNW33N carrying the β-(1,3-1,4) glucanase

(lichenase) gene of Streptococcus bovis genome was transferred to Bacillus subtilis

RSKK246 (CMCase+) strain by electroporation. Subsequently, electrotransformation was

performed on LB-agar plates containing lichenan and enzymatic activity regions of

recombinant B. subtilis RSKK246 colonies were observed by staining with Congo red. In

addition, the DNA from the recombinant plasmid pNW33N+Lichenase (pNW33NLic) was

cut on both the BamHI and HindIII endonucleases and observed on the lichenase gene

(1800 bp) agarose gel. On the other hand, the protein band corresponding to 26 kDa of

the recombinant enzyme was observed by zymogram analysis. These results indicate

that the β-(1,3-1,4) glucanase gene has been successfully expressed to the B. subtilis

strain RSKK246