SERINE PROTEASE DegP CHROMOHALOBACTER

0 downloads 0 Views 55KB Size Report
Keyword: Serine protease, DegP, halophilic bacteria.1. 1 Seminar Nasional Bioteknologi, “Bioteknologi untuk mendukung kualitas hidup. Bangsa. 2014.
SERINE  PROTEASE  DegP  CHROMOHALOBACTER  SALEXIGENS-­‐BKL-­‐5:     CLONING  AND  EXPRESSION       M.  Saifur  Rohman1,2,  Dewi  Fitriani2,  Irfan  D.  Prijambada1,2   1Laboratory  of  Agricultural  Microbiology,  Faculty  of  Agriculture,  Universitas   Gadjah  Mada;  2Graduate  Program  in  Biotechnology,  Universitas  Gadjah  Mada,   Yogyakarta-­‐INDONESIA       Abstract     Serine  protease  DegP  plays  an  important  role  in  protein  quality  control.    To  cope   its   important   role,   DegP   carries   out   dual   unique   functions   that   seem   to   be   opposing   to   each   other.   DegP   conveys   chaperon   and   protease   functions   in   a   single   entity.     The   expression   of   DegP   is   upregulated   in   response   to   various   environmental   stress   conditions,   such   as   high   temperature,   high   salt   concentration,   low   and   high   pH.   Halophilic   bacterium,   a   group   of   bacteria   that   able  to  grow  in  such  high  salt  concentration,  has  their  own  mechanism  to  protect   from  osmotic  stress.  The  common  mechanism  is  by  accumulation  of  compatible   solute   such   as   highly   water   soluble   sugar,   alcohols,   amino   acids,   betaines,   ectoines  or  their  derivatives.  In  regard  to  the  DegP,  it  is  not  known  whether  this   protein   is   involved   in   the   salt   tolerance   mechanism   of   halophilic   bacterium.   In   order   to   understand   of   the   role   of   DegP,   it   is   necessary   to   clone   and   overexpress   the   DegP   gene   from   halophilic   bacterium.   Therefore   the   objective   of   this   work   is   to  clone  and  overexpress  the  DegP  gene  from  Chromohalobacter  salexigens-­‐BKL-­‐ 5,  a  halophilic  bacterium  that  was  previously  isolated  from  Bledug  Kuwu  in  our   laboratory.   In   this   report,   we   have   successfully   cloned   and   over   expressed   the   DegP   gene   from   Chromohalobacter   salexignes-­‐BKL-­‐5   in   E.   coli   BL-­‐ 21(DE3)CodonPlus.  Analysis  of  the  DegP  encoding  gene  indicated  that  the  gene   consist   of   1356   bp   length   and   the   overexpressed   recombinant   protein   had   molecular  weight  of  49  kDa.  Solubility  check  analysis  indicated  that  the  protein   was   overexpressed   in   soluble   form.   Bioinformatics   analysis   showed   that   the   proteins  has  similar  characteristic  as  halophilic  proteins.         Keyword:  Serine  protease,  DegP,  halophilic  bacteria.1        

                                                                                                                1  Seminar  Nasional  Bioteknologi,  “Bioteknologi  untuk  mendukung  kualitas  hidup   Bangsa.  2014.  Sekolah  Pascasarjana  UGM