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Site-Directed Mutagenesis of Glu-269 L-Arabinose Isomerase from Geobacillus stearothermophilus Isolated from Tanjung Api Poso, Indonesia Fitriani, Dewi Fitriani; Wulandari, Puspita Suci; Saksono, Budi
ANNALES BOGORIENSES Vol 14, No 2 (2010): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (600.974 KB) | DOI: 10.1234/55

Abstract

Industrializing of tagatose requires enzymes that meet to industrial need such as thermophile, slightly acidic and metal independent.  Previously,  we  cloned,  sequenced  and  expressed  L-arabinose  isomerase  from  Geobacillus stearothermophilus  isolated  from Tanjung  Api, Poso, Indonesia. Based on DNA alignment analysis, the gene had  high  homology  with  those  of  G.  stearothermophilus  T6  (Gene  Bank  Acc  No:  AAD45718)  which  has optimum  activity  at  high  temperature  and  alkaline  condition.  In  this  p aper,  we  described  site-directed mutagenesis approach to mutate Glu-269 (Q269) to Lys-269 (K269) to decrease the optimum pH of the strain. Sequencing  result  showed  that  mutagenesis  had  been  successful  to  mutate  amino  acid  at  position  269  from glutamine (Q) into lysine (K). Expression of mutant Q269 showed protein with molecular mass ~56 kDa. Keywords: site-directed mutagenesis, Q269K, L-arabinose isomerase, Geobacillus stearothermophilus
Non Invasive Detection of Dengue Viruses from Saliva: In vitro Study Saksono, Budi
ANNALES BOGORIENSES Vol 22, No 2 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (214.71 KB) | DOI: 10.14203/ann.bogor.2018.v22.n2.75-80

Abstract

      In the previous paper, we had succeeded in developing an early detection system of dengue viruses using Sugar liganded Gold Nano Particle (SGNP) only from 6 μL serum. It has been reported that dengue virus is also detected in the saliva and urine of the patient. The evidences lead to the possibility of developing non-invasive methods of dengue virus detection. In this in vitro study, we evaluated the utility of SGNP to capture and concentrate dengue virion in 10% saliva solution. The results showed that dengue virion was successfully detected in 10% of saliva solution. Analysis of virion stability during storage showed that virions in salivary samples were stable up to 3 days at temperature wherease the RNA has significantly degraded. Although still a preliminary study, the data obtained show the prospect of SGNP as a non-invasive dengue virus detection method, as well as the development of POC (Point of Care) method. Clinical trials using saliva from dengue viruses infected patients need to be done to prove the effectiveness of the SGNP method.