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Journal : UNEJ e-Proceeding

The Potency of Protein Extracts from Candida albicansas Bioreceptor on Immunosensor for Diagnosis of Candidiasis ., Masfufatun; Kumala, Noer; Baktir, Afaf
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

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Abstract

Currently diagnosis of candidiasis still usingthe traditional standard blood culture method. The traditional method were less sensitive andtime consuming. The purpose of this research were to develope the more sensitive immunosensor based method, and to examine the potency of C. albicans protein extract as bioreceptor to  detect  C. albicans and its biofilm in the blood of candidiasis patients.The research methods include: (1) preparation of  digestive gland liquid of snail (Achatina fulica); (2) extraction of protein from C. albicans  through  enzimatis and mechanic methodsand (3) analyzing the protein extract as bioreseptor through immunodot assay.The research results showed that the snail enzymes has protein content 1.35 mg/ml and  specific activity 1.96 unit/mg. The snail enzyme hydrolyzed  the cell wall of C. albicans  with and without sonication, producedplanktonic extracell protein extract (PEP) and biofilmextracell proteinextract (PEB), planktonic intracell protein extract (PIP), and biofilmintracell protein extract (PIB), withprotein content 1.44; 1.29; 1.29 and 1.21 mg/ml respectively. Thebiofilm intracell protein (BIP) showed antigenic property towardantibody anti-Candida (positive control),giving red spot on imunodot assay. Immunodot assay can distinguishnegative control serum (health man) and positive Candidiasis control by using antigen 1 mg/ml and50ml serum. Keywords: C. albicans, candidiasis, biofilm, immunodot assay
Exploration of Lipase Enzyme from Soil through Metagenomic Approach Sumarsih, Sri; Baktir, Afaf; Andina, Budi Putri Ayu
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

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Abstract

This research aimed to explore lipase enzyme from soil through metagenomic approach. The DNA was extracted directly from the topsoil and river sediment by SDS-based lysis method. The soil genomic DNA was sheared into approximately 40-kb fragments The construction of genomic library was carried out by cloning the fragment DNA into  fosmid pCC2FOS vector and host E. coli EPI300-T1R accordance with the protocols of CopyControl™ Fosmid production library kit. Screening of lipase-producing clones was performed based on its lipolytic activity using Luria-Bertani agar plate 12.5 µg/ ml chloramphenicol, 10 µg/ml Rhodamine B and 1 % olive oil as a substrate. The assay plates were incubated at 550 C. The lipolytic activity was identified as an orange around colonies. There were 5 (five) clones of 32 screened-clones that expressed lipolytic activity. The next research will characterize the enzyme and determine the sequence of the enzyme-encoding gene. Keywords: Lipase, soil, metagenomic, fosmid vector, pCC2FOS, E. coli EPI300-T1R