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All Journal JURNAL EMBA : JURNAL RISET EKONOMI, MANAJEMEN, BISNIS DAN AKUNTANSI Mechatronics, Electrical Power, and Vehicular Technology Briliant: Jurnal Riset dan Konseptual ANNALES BOGORIENSES JURNAL PENDIDIKAN TAMBUSAI Jurnal Basicedu Jurnal Yustisiabel Journal of Public Health Innovation (JPHI) International Journal of Educational Review, Law And Social Sciences (IJERLAS) Jurnal Administrasi Bisnis Makara Journal of Technology Jurnal Basicedu Indonesian Journal of Community Empowerment Jurnal Indonesia Sosial Teknologi Jurnal Kesehatan Bidkemas Bata Ilyas Educational Management Review Annales Bogorienses
Dian Andriani, Dian
Interdisciplinary Program of Graduate School for Bioenergy and Biomaterials, Chonnam National University,Gwangju 500-757
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Journal : ANNALES BOGORIENSES

 1 
Generation of mCherryBody: an Anti-Transferrin Receptor Antibody Variable Fragment Linked by The Fluorescent Protein mCherry Kusharyoto, Wien; Andriani, Dian; Handayani, Ira
ANNALES BOGORIENSES Vol 20, No 2 (2016): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v20i2.233

Abstract

A facile generation of a recombinant antibody fragment with intrinsic fluorescent properties of the monomeric fluorescent protein mCherry is described. The so-called mCherryBody was designed based on the structure model of the variable fragment of anti-transferrin receptor antibody LUCA31 and the X-ray crystallographic structure of the protein mCherry. mCherryBody was constructed to retain optimal spatial geometry between the C- and N-termini of the antibody light-chain (VL) and heavy-chain (VH) by mimicking the domains interface pairing in antibody Fab fragments and incorporation of the fluorescent protein mCherry as a bridging scaffold. The gene encoding the chimeric protein was cloned into the pJExpress414 expression vector, expressed and secreted into the periplasm of Escherichia coli NiCo21(DE3) for assembly and disulphide bond formation. Based on its amino acid sequence, mCherryBody was predicted to have a molecular weight of 51.46 kDa. The modular assembly used in the generation of mCherryBody may permit the interchange of binding sites and of fluorescent proteins to create robust panels of coloured antibody fragments. Thus, the mCherryBody platform facilitates rapid generation of colored single-chain variable fragment (scFv) chimeras that could be used for screening of antibodies against cell surface markers or receptors.
Screening for Natural Producers Capable of Producing 1,3-Propanediol from Glycerol Andriani, Dian; Kusharyoto, Wien; Prasetya, Bambang; Wilke, Thomas; Vorlop, Klaus Dieter
ANNALES BOGORIENSES Vol 14, No 1 (2010): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/43

Abstract

Glycerol is a renewable resource found as the main  by-product in the transesterification of triglycerides and fat saponification. Due to the increased production of plant oils, especially palm  oil  in developing countries, and their larger use  by the oleochemical industry, glycerol surpluses are on the world market and this may result in a desrease in glycerol  price. As a consequence, biotechnological processes  have been developed to convert this substrate  into  value-added  products,  such  as  1,3-propanediol  (1,3-PD).  The  microbial  production  of  1,3-PD could  be  competitive  to  chemical  routes  assuming  that  it  is  based  on  cheap  raw  material  and  an  optimized process.  In  the  screening  for  1,3  PD–producing  bacteria,  raw  glycerol  as  by-product  from  rapeseed  oil processing unit  was  used  as  a  carbon  source  compared  with  commercial  glycerol.  By  using  increasing concentration of  both  glycerols  from  50  to  150  g/l,  two  potential  bacteria  were  obtained  from  soil  samples.BMP 1 was obtained from an enrichment culture using 50 g/l commercial glycerol, while BMR-1 was obtained from  an enrichment culture using 100 g/l raw glycerol. The highest conversion yield obtained using the isolateBMP-1 was around 0.62 g 1,3-PD formed per  mol glycerol consumed, and 0.73  mol 1,3-PD  formed per  molgycerol using the isolate BMR-1. No bacteria were obtained from cultures using 150 g/l commercial and rawgycerol, respectively, which indicated that higher concentration of glycerol has inhibition effect.   Keywords: 1,3-propanediol, enrichment culture, glycerol, palm oil, screening
Expression of an Anti-Transferrin Receptor Antibody SCFV Fragment in Escherichia coli Using A L-Rhamnose-Based Tightly Regulated Promoter System Andriani, Dian; Handayani, Ira; Kusharyoto, Wien
ANNALES BOGORIENSES Vol 16, No 2 (2012): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3611.759 KB) | DOI: 10.1234/63

Abstract

Transferrin receptor (TfR) levels are elevated in various types of cancer cells and correlate with the aggressive or proliferative ability of tumor cells. Therefore, TfR levels are considered useful as a prognostic tumor marker, and TfR is a potential target for drug delivery in therapy of malignant cells. In such kind of targeted delivery system, antibody fragments are frequently used as targeting moiety. Here, we report the generation of an anti-TfR single-chain antibody variable (scFv). The cDNA encoding the variable of heavy and light chain domains of the scFv antibody fragment was derived from the anti-TfR monoclonal antibody of LUCA31. The gene encoding the anti-TfR scFv fragment was codon, optimized for expression in Escherichia coli, subsequently synthesized, and cloned into the pJExpress-804 Rhamex vector. The expression vector utilizes the E. colirhaB promoter and corresponds to the regulatory genes, and is tightly regulated by the presence of L-L-rhamnose. It is also tightly regulated in the absence of L-L-rhamnose by the addition of D-glucose The His6-tagged anti-TfR scFv fragment was expressed in E. coliNiCo21 and purified by means of immobilized metal chelate affinity chromatography on TALON™ matrix. In SDS-PAGE, a single band corresponding to a molecular mass of approximately 30 kDa was observed whether it corresponded to the predicted molecular mass based on the amino acid sequence.Keywords: antibody fragment, L-L-rhamnose, Rhamex vector, scFv, transferrin receptor
Screening and Optimization of Cellulase Production of Bacillus subtilis TD6 Isolated from Takifugu rubripes Fish Park, Don Hee; Andriani, Dian
ANNALES BOGORIENSES Vol 14, No 1 (2010): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/40

Abstract

Cellulase enzymes have attracted considerable attention in recent years due to their great biotechnological and industrial potential.  Cellulase enzymes provide a key opportunity for achieving tremendous benefits of biomass utilization through  the  bioconversion  of  the  most  abundant  cellulosic  material  into  the  simplest  carbohydrate monomer,  glucose.  Nowadays,  the  sources  of  cellulase-producing  bacteria  have  been  broadened  into  thepresence  of  symbiotic  bacteria  in  herbivorous  animal  and  also  from  marine.  Takifugu  rubripes  or  known  as puffer fish is a unique poisonous vertebrate  but nevertheless is considered a delicacy  in Korea. The diet of the puffer fish includes mostly algae. This dietary habit considers Puffer fish as host of cellulase-producing bacteria, especialy on its gut.  In the present study an attempt has been    made to search for the cellulolytic bacteria in the gut of Takifugu rubripes. Fifty five microorganisms have been isolated using 1% (w/v) Carboxymethyl cellulose (CMC) as substante Congo red  dye  test  and  DNS  method  were  then  used  for  screening  the  extracellular celuase activity of the strains. Among them, TD6 strain has shown the highest performance in term of cellulose activity. In order to evaluate the optimum culture condition of the isolate TD6 for cellulase production, the strain was grown at various temperatures, pH, carbon  sources, and  nitrogen sources.   Under optimum  condition, the maximum specific activity of 2.13 U/mg protein was achieved after growth the strain with 1.5% CMC at 45ºC PH 6 for 3 days, respectively Based on 16S rRNA gene analysis it is proposed that the strain was identified asBacillus subtilis.
Co-Authors Alif Mudiono Bakri , Muhammad Bambang Prasetya Bambang Suteng Sulasmono Burhanuddin Basri Changshin Sunwoo, Changshin Christoffel Kojo Cukanda, Cucu Dandan Haryono Darussalam, Rudi Don-Hee Park, Don-Hee Dwijayati, Gita Nofieka Edita Martini, Edita Erna Safariyah Eunice Widyanti Setyaningtyas Fadjriani, Lia Fahroji, Fahroji Fransiska Novita Eleanora Hana, Ju Handayani, Ira Handayani, Ira HARYADI, ARDI MULYANA Hendarsyah, Hendarsyah Hendra N. Tawas Indradewa, Rhian Julianto, Agus Tedi Kang, Kyeong Eop Kurnia, Aji Kusharyoto, Wien Kusharyoto, Wien Kusumah, Riyana Kyeong Kang, Kyeong Lina Herlina Mardiana, Mira Milanie, Feby Mulyani, Nenti Antalanti Mulyasari, Iis Negara, Chandra Perwira Park, Don Hee Park, Don Hee Prariatna, Frisca Puji Muniarty Rohmani, Tina Yunia Rony, Zahara Tussoleha Rusli, Damay Saifannur, Andri sanusi, Anwar Sari, Roza Puspita Setiabudi, Dedi Sriramulu GobiKrishnan, Sriramulu Surn-Teh Bark, Surn-Teh Suroyo Suroyo Syamsul Bahri Tinton Dwi Atmaja Vivi Novianti Vorlop, Klaus Dieter Vorlop, Klaus Dieter Washliati, Laily Wilke, Thomas Willke, Thomas Yunie, Chanty