cover
Filter by Year
Articles by issue : Vol 11, No 1 (2017): March 2017
5
Articles
Cloning of Synthetic Lipase Gene from Rhizomucor miehei with Original Signal Peptide in Pichia pastoris

CAHYANI, MARTHA EKA, HELIANTI, IS, NURHAYATI, NIKNIK, ABINAWANTO, ABINAWANTO

Microbiology Indonesia Vol 11, No 1 (2017): March 2017
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (845.107 KB)

Abstract

Lipases (EC 3.1.1.3) are classified as hydrolases that hydrolyze lipids. These enzymes have potential application in biotechnology and industrial process. In previous study we have cloned the synthetic Rhizomucor miehei lipase gene using the vector pUC57 in Escherichia coli DH5α, but only found the very low enzymes activity. This study aimed to clone Rhizomucor miehei synthetic lipase gene into Pichia pastoris expression plasmid for lipase expression with the original signal peptide. A DNA fragment with the original signal peptide had been obtained by PCR, cut by Xho I and Xba I and then ligated into pPICZα A linearized with the same enzymes. The mixture of ligation, then was transformed into Escherichia coli DH5α. Zeocin-resistant transformants were selected and contained plasmid was analyzed by restriction enzymes analyses, PCR, and DNA sequenced. As the result, a Rhizomucor miehei lipase gene (RMlip) with the size of 1132 bp was successfully cloned to pPICZα A. The recombinant plasmid with the correct DNA sequence was transformed into Pichia pastoris X33. Cultivation of recombinant P. pastoris was carried out with the addition of 1.5% methanol every day with appropriate aeration. The recombinant lipase produced by Pichia pastoris X33 containing RMlip oin its chromosomal DNA had optimal temperature and pH 30 C and 9.0, respectively.

ITA REGISTRATION FORM AND BACK COVER

Helianti, Is ( 1. BPPT 2. Indonesian Society for Microbiology )

Microbiology Indonesia Vol 11, No 1 (2017): March 2017
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (1340.746 KB)

Abstract

Analysis of Human Immune Response against Salivary Glands Protein Extract of Anopheles sundaicus. L in Malaria Endemic Area

NURYADY, MOHAMMAD MIRZA ( Jember University ) , UTOMO, SUGENG SEYO, ARMIYANTI, YUNITA, WIDJAJATI, SRI MUMPUNI WAHYU, SENJARINI, KARTIKA

Microbiology Indonesia Vol 11, No 1 (2017): March 2017
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (566.154 KB)

Abstract

Malaria is an infectious disease caused by Plasmodium, which is transmitted by Anopheles mosquitoes as vectors. Malaria transmission begins when an infected mosquito takes blood meal from healthy human. Mosquitoes will release parasite and components of saliva into the hosts body. Saliva contains components (proteins) that affect the hosts hemostasis and immune respose, such as vasomodulator and immunomodulators. Imunomudulator could act as immunosuppressive factors that can suppress nonspecific immune system of the host and modulate the change of T helper 1 (Th1) toward T helper 2 (Th2) response, which is advantageous for malaria parasite to infect human host. This research wanted to evaluate human immune respons in endemic area against salivary gland protein extract (SGPE) from its major malaria vector i.e. Anopheles sundaicus (An. sundaicus). Analysis of human immune response was conducted quantitatively by ELISA (Enzyme Link Immunosorbend Assay) towards IgG from human sera samples after cross reacted with SGPE. The results showed that exposures to An. sundaicus were able to induce high levels of IgG. IgG anti salivary proteins of An. sundaicus is higher than the levels of IgG anti salivary proteins of Ae. aegypti. Furthermore, the age group 11-40 years with the highest bites probability, had the highest IgG levels compared to other age groups.

Construction and Expression of Single Recombinant Peptide Surfactant for EOR Application

SARI, CUT NANDA, USMAN, USMAN, ROHMAT, RIESA KW, HERLINA, LENI, SULIANDRI, KEN SAWITRI, KRISTIAWAN, ONIE, DWIYANTARI, DWIYANTARI, KRISTIANTI, TATI, SUHANDONO, SONY

Microbiology Indonesia Vol 11, No 1 (2017): March 2017
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (985.809 KB)

Abstract

Surfactant is generally synthetic chemical, which is effective and reliable. However, the chemicals usually did not degraded easily in the environment and could cause damage to the environment. The other possible alternative to produce surfactant is using genetic engineering in order to produce peptide based surfactant. In this research, peptide surfactant was produced using a gene construct which was created using overlapped polymerase chain reaction method (OE-PCR). PAGE analysis shows that single surfactant peptide construction can be expressed by induction of IPTG 1 mM and after at least twice sonication. This research proves that both two constructions have been successfully expressed by producing peptide in expected size (approximately 15 kDa).

Studies for IAA (Indole-3-Acetic Acid) Production by Isolates H6 with Nitric Acid Mutation

PUTRIE, RAHAYU FITRIANI WANGSA, WIDOWATI, TIWIT, SUKIMAN, HARMASTINI

Microbiology Indonesia Vol 11, No 1 (2017): March 2017
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (507.372 KB)

Abstract

Nitric acid mutations are known could be used for strain improvement. This research aimed to studies IAA production by nitric acid mutan were compared with wild type. Mutation were conducted with some different treatment time such as 0, 30, 60, 90 and 120 min subsequently it were measured for IAA production. Isolate H6 as -1wild type isolates were also molecularly identified. The wild strain exhibited 53.83 µg mL of IAA while the -1 -1. nitric acid mutan within a range 77.39 µg mL to 95.70 µg mL Isolates H6.60 exhibited the highest IAA -1 production which 39.87 µg mL higher were compared with wild-type. Based on 16S rRNA gene analysis, isolate H6 belonged to Lysobacter sp. ES2-22.