Ariyanti Oetari

4,6-α-Glucanotransferase, a Novel Enzyme That Structurally and Functionally Provides an Evolutionary Link between Glycoside Hydrolase Enzyme Families 13 and 70

ABSTRACT Lactobacillus reuteri 121 uses the glucosyltransferase A (GTFA) enzyme to convert sucrose into large amounts of the α-d-glucan reuteran, an exopolysaccharide. Upstream of gtfA lies another putative glucansucrase gene, designated gtfB. Previously, we have shown that the purified recombinant GTFB protein/enzyme is inactive with sucrose. Various homologs of gtfB are present in other Lactobacillus strains, including the L. reuteri type strain, DSM 20016, the genome sequence of which is available. Here we report that GTFB is a novel α-glucanotransferase enzyme with disproportionating (cleaving α1→4 and synthesizing α1→6 and α1→4 glycosidic linkages) and α1→6 polymerizing types of activity on maltotetraose and larger maltooligosaccharide substrates (in short, it is a 4,6-α-glucanotransferase). Characterization of the types of compounds synthesized from maltoheptaose by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS), methylation analysis, and 1-dimensional 1H nuclear magnetic resonance (NMR) spectroscopy revealed that only linear products were made and that with increasing degrees of polymerization (DP), more α1→6 glycosidic linkages were introduced into the final products, ranging from 18% in the incubation mixture to 33% in an enriched fraction. In view of its primary structure, GTFB clearly is a member of the glycoside hydrolase 70 (GH70) family, comprising enzymes with a permuted (β/α)8 barrel that use sucrose to synthesize α-d-glucan polymers. The GTFB enzyme reaction and product specificities, however, are novel for the GH70 family, resembling those of the GH13 α-amylase type of enzymes in using maltooligosaccharides as substrates but differing in introducing a series of α1→6 glycosidic linkages into linear oligosaccharide products. We conclude that GTFB represents a novel evolutionary intermediate between the GH13 and GH70 enzyme families, and we speculate about its origin.

Detection of fungi from old Chinese manuscripts in Central Library Universitas Indonesia

Old manuscripts, as organic materials, can be subjected to deterioration by fungi [1]. Fungal species (over 200) are the main cause of damage to objects of cultural heritage made of or supported on paper [2]. Fungi from old manuscripts can be identified to species level by analysis of Internal Transcribed Spacer (ITS) region of rDNA [3]. Old Chinese manuscripts in Indonesia are one of historical evidences of close relationship between Indonesia and China for hundred years. From our preliminary survey, the collection of old Chinese manuscripts in Central Library Universitas Indonesia was deteriorated by fungi. Information about fungal species from old Chinese manuscripts has not been reported yet, both in Indonesia and China. Our study is aimed to detect the growth of fungi on old Chinese manuscripts, and to isolate and identify the fungi that deteriorates the manuscripts in Central Library Universitas Indonesia. Twelve samples of old Chinese manuscripts deteriorated by fungi shows spore formation and brown spot on their paper surface. Forty one fungal isolates are collected from seven manuscripts, and many of them (28 isolates from 41) are xerophiles. Fifteen isolates were identified by molecular approach using ITS5. They were identified as Anthostomella sp., Aspergillus aculeatus, Aspergillus insuetus, Chaetosartorya stromatoides, Cladosporium colocasiae, Cladosporium columbiae, Cladosporium pini-ponderosa, Flavomyces fulophazii, Fusarium equiseti, Fusarium sp., and Pseudozyma sp. Fifteen fungal isolates could be identified to genus level based on morphological characters as follows: Aspergillus (10 isolates), Curvularia (1 isolate), and Penicillium (4 isolates). Some isolates could not be identified to the genus level since they do not produce asexual/sexual spores (mycelia sterilia). Further identification to species level will be carried out by molecular approach. Most of the isolated fungi from old Chinese manuscripts were xerophiles and able to use Chinese paper as a substrate. This study is the first report on the diversity of fungi from old Chinese manuscripts from Indonesia.Old manuscripts, as organic materials, can be subjected to deterioration by fungi [1]. Fungal species (over 200) are the main cause of damage to objects of cultural heritage made of or supported on paper [2]. Fungi from old manuscripts can be identified to species level by analysis of Internal Transcribed Spacer (ITS) region of rDNA [3]. Old Chinese manuscripts in Indonesia are one of historical evidences of close relationship between Indonesia and China for hundred years. From our preliminary survey, the collection of old Chinese manuscripts in Central Library Universitas Indonesia was deteriorated by fungi. Information about fungal species from old Chinese manuscripts has not been reported yet, both in Indonesia and China. Our study is aimed to detect the growth of fungi on old Chinese manuscripts, and to isolate and identify the fungi that deteriorates the manuscripts in Central Library Universitas Indonesia. Twelve samples of old Chinese manuscripts deteriorated by fungi shows spore formation and brow...

Diversity of fungi from old Dluwang manuscripts from Mertasinga, Cirebon

In the 17th century, Javanese handwritten manuscripts were written on dluwang. The material was made from the bark of the paper mulberry (Broussonetia papyrifera Vent.) [1]. Microbial consortia, however, may inhabit ancient manuscripts. Cellulolytic organisms can exploit the paper as substrate [2]. Furthermore, some fungi which are able to grow in dry substrate (xerophilic character) are associated with deterioration of manuscripts [3]. A local collector from Mertasinga, Cirebon, Indonesia, preserves old dluwang manuscripts according to traditional Javanese custom. In our previous study, twenty three fungal isolates were obtained from the teak wardrobe which contained old dluwang manuscripts. The fungal isolates are deposited in the Universitas Indonesia Culture Collection (UICC). The aim of our study was to isolate and identify fungi from old dluwang manuscripts and to identify fungal isolates obtained from previous study. Ten old dluwang manuscripts showed fungal growth, characterized by spore-formation...

Paenibacillus cisolokensis sp. nov., isolated from litter of a geyser

A Gram-stain-positive, endospore-forming, aerobic and thermophilic bacterium, designated strain LC2-13AT, was isolated from Cisolok geyser, West Java, Indonesia, at 50 °C. The isolate was rod-shaped and motile by means of peritrichous flagella. The major cellular fatty acids were iso-C16 : 0, C16 : 0 and anteiso-C15 : 0 and the major quinone was menaquinone 7. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The genomic DNA G+C content was 56.6 mol% and the major diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain LC2-13AT is related most closely to Paenibacillus kobensis DSM 10249T (94.86 % similarity), Paenibacillus tarimensis SA-7-6T (94.77 %) and Paenibacillus barengoltzii SAFN-016T (94.77 %). On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, strain LC2-13AT is affiliated to the genus Paenibacillus, but could be distinguished from recognized species of this genus. A novel species with the name Paenibacillus cisolokensis sp. nov. is thus proposed. The type strain is LC2-13AT (=UICC B-42T=NRRL B-65368T=DSM 101873T).

Polyphasic identification of a thermophilic bacterium from geyser of Cisolok, Indonesia

A thermophilic bacterium strain UICC B-76 was isolated from litter in geyser of Cisolok, West Java, Indonesia by using The International Streptomyces Project (ISP1) medium. The strain UICC B-76 was able to hydrolyze various substrates such as casein, cellulose, gelatin, starch, and xylan, at 50 °C, and decolorize Remazol Brilliant Blue R (RBBR) at 50 °C, an indicator for ligninolytic degradation ability. These abilities of strain UICC B-76 suggest its potential for industrial applications. The aim of this study was to identify strain UICC B-76 based on phenotypic and genotypic characterizations. Strain UICC B-76 was Gram-positive, spore-forming, aerobic and facultative anaerobic, and rod-shaped with flagella. The strain grew at 30 °C to 60 °C (optimum 50 °C) and pH 7.0 but can not withstand temperature of 70 °C as well as pH 4.0 and 9.0. Strain UICC B-76 could not grow in more than 1% NaCl. This strain has menaquinone 7 (MK-7) as the major quinone system; cell wall fatty acids consisted of Iso-C15:0 (68.5...

The Effect of Pollen Substitutes on the Productivity of Apis cerana in Indonesia

The aims of this study were to examine the preferences and productivity of A. cerana colonies when fed on a variety of pollen substitutes.

PENGEMBANGAN DATABASE MIKROORGANISME INDIGENOS INDONESIA

Abstract Development of Database for Indigenous Indonesian Microorganisms. The objective of the research is to create anddevelop a database of indigenous Indonesian microorganisms based at the University of Indonesia. Development of thedatabase of indigenous Indonesian microorganisms was carried out in several stages, i.e. data identification, databasedesign, programming, data entry, testing and debugging, and repairing and maintenance. Development of the databaseutilized the licensed software of General Public License (GPL), which include Linux RedHat 9.0 (operating system),Apache ver. 2.20 (web server), MySQL ver. 4.2 (database server), and PHP ver. 4.3 (web interface programminglanguage). The result of this research is a database named UI Bioinfo which has the following facilities: online catalogsearch for UICC (University of Indonesia Culture Collection) strains collection and sequence homology search utilitythrough BLAST (Basic Local Alignment Search Tool). Integrated information on strains collection was first carried outon the yeast collection. At present, UI Bioinfo contains information for 297 strains that includes isolation data,morphological descriptions, physiology-biochemical characteristics, and images. Moreover it also contains sequencedata from the large subunit (LSU) ribosomal RNA gene and the internal transcribed spacer (ITS) regions. UI Bioinfocan be accessed from the following site: http://152.118.162.250/bio/. Future development will be addition of data fromthe other collections in UICC.Keywords:Bioinformatics, database, Indonesia, microorganisms