Various host systems have been employed to increase the yield of recombinant proteins. However, some recombinant proteins were successfully produced at high yields but with no functional activities. To achieve both high protein yield and high activities, molecular biological strategies have been continuously developed. This work describes the effect of signal peptide (SP) and co-expression of molecular chaperones on the production of active recombinant protein in Escherichia coli. Extracellular enzymes from Bacillus subtilis, including β-1,4-xylanase, β-1,4-glucanase, and β-mannanase constructed with and without their signal peptides and intracellular enzymes from Pseudomonas stutzeri ST201, including benzoylformate decarboxylase (BFDC), benzaldehyde dehydrogenase (BADH), and d-phenylglycine aminotransferase (d-PhgAT) were cloned and overexpressed in E. coli BL21(DE3). Co-expression of molecular chaperones with all enzymes studied was also investigated. Yields of β-1,4-xylanase (Xyn), β-1,4-glucanase (Cel), and β-mannanase (Man), when constructed without their N-terminal signal peptides, increased 1112.61-, 1.75-, and 1.12-fold, respectively, compared to those of spXyn, spCel, and spMan, when constructed with their signal peptides. For the natural intracellular enzymes, the chaperones, GroEL-GroES complex, increased yields of active BFDC, BADH, and d-PhgAT, up to 1.31-, 4.94- and 37.93-fold, respectively, and also increased yields of Man and Xyn up to 1.53- and 3.46-fold, respectively, while other chaperones including DnaK-DnaJ-GrpE and Trigger factor (Tf) showed variable effects with these enzymes. This study successfully cloned and overexpressed extracellular and intracellular enzymes in E. coli BL21(DE3). When the signal peptide regions of the secretory enzymes were removed, yields of active enzymes were higher than those with intact signal peptides. In addition, a higher yield of active enzymes was obtained, in general, when these enzymes were co-expressed with appropriate chaperones. Therefore, E. coli can produce cytoplasmic and secretory enzymes effectively if only the enzyme coding sequence without its signal peptide is used and appropriate chaperones are co-expressed to assist in correct folding.

Suhardi, Summpunn P, Duangjinda M, Wuthisuthimethavee S. 2020. Phenotypic diversity characterization of Kalang and Thale Noi Buffalo (Bubalus bubalis) in Indonesia and Thailand: Perspectives for the buffalo breeding development. Biodiversitas 21: 5128-5137. Kalang (KBuf) and Thale Noi buffaloes (TBuf) are swamp buffalo breeds important for biodiversity resources and permeate culture, religion, meat supply, and livelihood in Indonesia and Thailand. This study aimed to analyze the phenotypic characterization within breeds of KBuf/TBuf using quantitative and qualitative performance to define diversity. A total of 240 buffaloes from North (NK), East (EK), South Kalimantan (SK), Indonesia, and Phatthalung (PT), Thailand were observed. Phenotypic characterization showed female TBuf had the highest quantitative variable values with significant difference (P0.05) and females from EK, SK, and PT showed a close relationship (P>0.05). The cumulative index results demonstrated KBuf/TBuf as very good meat-producing livestock. In conclusion, variations in qualitative and quantitative variables indicate phenotypic diversity in KBuf/TBuf which is a basis to perform selection and breeding development through pedigree recording, reproductive efficiency, cross-breeding, and genetic modification to produce high-quality buffalo breeding stock. KBuf/TBuf can provide high-quality red meat through improved genetic quality, productivity, and economic management towards modern buffalo breeding development.

Kalang (KBuf), Krayan (KrBuf), and Thale Noi buffaloes (TBuf) are swamp buffalo genetic resources in Indonesia and Thailand. The maternally inherited mitochondrial DNA (mtDNA), particularly D-loop region is an important material for phylogenetic inference and analyzing genetic diversity. Therefore, the objectives of the present study were to evaluate genetic diversity and to reconstruct the phylogenetic tree within buffalo breeds in Kalimantan, Indonesia, and Phatthalung, Thailand using mtDNA D-loop sequences. A total of one hundred forty buffaloes (70 males and 70 females) were observed including 40 buffaloes from North (NK), 40 from East (EK), and 40 from South Kalimantan (SK) provinces Indonesia and 20 from Phatthalung (PT) province, Thailand. DNA samples were isolated from buffalo tail hairs. DNA sequences were manually assembled using BioEdit program with consideration of gaps and ambiguous sequences. The phylogenetic tree of buffalo was generated by PHYLIP software. The observed variables included haplotype diversity, genetic distance, and genetic tree. The 956 bp of amplified mtDNA D-loop fragment presented a total of 24 haplotypes with several mutations that included transitions (293), transversions (60), deletions (15), and insertions (20). The neighbor-joining tree using the Kimura 2 parameter model demonstrated two local buffalo clusters among buffalo from Kalimantan and Thailand with four buffalo relationship patterns observed from buffaloes in Kalimantan Island (KBuf and KrBuf), Indonesia. The Results of the present study demonstrated that the buffaloes sequence analysis revealed relatively high diversity and is a good basis to perform selection and modern buffalo breeding development.

Swamp buffalo (Bubalus bubalis) is one of the Indonesian germplasm that adapts well to Borneo Island. This study aimed to determine the genetic diversity, phylogenetic tree, and phylogeographic structure of Bornean swamp buffalo based on control region (D-loop) mitochondrial DNA sequences. A total of 120 animals were collected from three populations in Indonesia, i.e., the North, East, and South of Kalimantan Provinces. Along 1140 bp of D-loop mtDNA gene of Bornean swamp buffaloes was amplified with a design primer of F: 5-CAA CAC CCA AAG CTG AAG TT-3’ and R: 5-CGC TCC TCT TAG TCT CGT TG-3’. Therefore, the forward and reverse sequencing was performed to visualize the full length of D-loop mtDNA gene sequence (1140 bp). Results showed that a total of 47 haplotypes were detected in the animal study, with haplotype and nucleotide diversities of 0.936 and 0.005, respectively. Consequently, two haplogroups were observed in the animal study, i.e., Haplogroup A (North) and Haplogroup B (East and South). Based on structure analysis, Bornean swamp buffaloes were comparable to filing swamp buffaloes of China based on structure. According to the analysis of molecular (AMOVA), the geographical component contributed over 56.44% of the total mtDNA sequence variations. In conclusion, it was discovered that the haplogroups of buffalo from the East and South populations were identical.

PENYULUHAN LITERASI DIGITAL MELALUI SOSIALISASI PADA ANAK-ANAK DI SDN 007 MUARA KAMAN

PENERAPAN ZERO WASTE MELALUI PELATIHAN DAUR ULANG SAMPAH ANORGANIK DENGAN MENGGUNAKAN METODE ECOBRICK