Drought stress is the primary constraint on plant-based food production, particularly paddy production. Several studies have examined plant resistance to heat and osmotic pressure. This study aimed to isolate bacteria with plant growth-promoting properties that could tolerate high temperatures and improve paddy growth during drought. Five isolates with a high optical density value (OD600) at 30% PEG 6000 (equivalent to -1.03MPa) and able to grow at high temperatures were identified based on the 16S rRNA gene sequences as Achromobacter spanius UKM UR10, Bacillus pumillus UKM UR11, Bacillus cereus UKM R66, and Bacillus altitudinis UKM RB11, which were isolated from the root, where as Bacillus sp. UKM S8 was isolated from rhizosphere soil. These isolates exhibited 1-amino cyclopropane-1-carboxylate (ACC) deaminase activities ranging between 1.01 and 1.12 mmol α-ketobutyrate mg-1 protein h-1, which degraded ACC to α-ketobutyrate and ammonia. Other plant growth promoters assessed include indole acetic acid (IAA; concentration between 9.69 and 13.15µg/ml) and phosphate solubilization (concentrations between 31.74 and 51.30 mg/l) production. Subsequently, the selected plant growth-promoting rhizobacteria (PGPR) were incorporated as a consortium and inoculated on paddy seeds, thus increasing total chlorophyll, proline, and soluble sugar content in paddy subjected to drought-stress conditions. Paddy yield components and performances, such as panicle number, spikelet number, dry grain weight, number of leaves, stalk length, and root length increased significantly. This isolated PGPR exhibits heat resistance, promotes plant growth, and can serve as an inoculant for paddy plants under drought conditions.
Microbial proteases constitute one of the most important groups of industrially relevant enzymes. Proline iminopeptidases (PIPs) that specifically release amino-terminal proline from peptides are of major interest for applications in food biotechnology. Proline iminopeptidase has been extensively characterised in bacteria and filamentous fungi. However, no similar reports exist for yeasts. In this study, a protease gene from Glaciozyma antarctica designated as GaPIP was cloned and overexpressed in Escherichia coli. Sequence analyses of the gene revealed a 960 bp open reading frame encoding a 319 amino acid protein (35,406 Da). The purified recombinant GaPIP showed a specific activity of 3561 Umg−1 towards L-proline-p-nitroanilide, confirming its identity as a proline iminopeptidase. GaPIP is a cold-active enzyme with an optimum activity of 30 °C at pH 7.0. The enzyme is stable between pH 7.0 and 8.0 and able to retain its activity at 10–30 °C. Although GaPIP is a serine protease, only 25% inhibition by the serine protease inhibitor, phenylmethanesulfonylfluoride (PMSF) was recorded. This enzyme is strongly inhibited by the presence of EDTA, suggesting that it is a metalloenzyme. The dimeric structure of GaPIP was determined at a resolution of 2.4 Å. To date, GaPIP is the first characterised PIP from yeasts and the structure of GaPIP is the first structure for PIP from eukaryotes.
Lichen is a symbiotic organism that exists as a single composite body consisting of a mycobiont (fungus) and a photobiont (algae or a cyanobacterium). Many lichen species are considered as extremophiles due to their tolerance to radiation, desiccation, temperature and pollution. However, not all lichen species are tolerant to harsh environmental conditions as several species are sensitive for example to nitrogen, sulphur, acidity, heavy metals, halogens (e.g. fluoride) and ozone. Thus, to better understand why some lichens can withstand exposure to pollutants as opposed to those that are susceptible, we focused on the lichen species of Dirinaria known for their wide distribution in the tropics, subtropics and pantropical, and moderate tolerance to air pollution. Their moderate tolerance to air pollution affords them to thrive in good air quality environments as well as polluted air environments. Lichen samples of Dirinaria sp., UKM-J1 and UKM-K1, were respectively collected from two areas with different levels of air quality based on Air Pollutant Index or API (with index pollutant criteria of PM10, carbon monoxide, ozone, nitrogen dioxide and sulfur dioxide) in the outskirt of Jerantut (UKM-J1), a rural area in the middle of Peninsular Malaysia and the township of Klang (UKM-K1), in a busy area of the Klang Valley, Malaysia. API was monitored throughout 2012-2013 whereby the sample collection site in Klang showed markedly higher concentrations of pollutants in all the index pollutant criteria as compared to that of Jerantut. We performed transcriptome sequencing using Illumina RNA-seq technology and de novo assembly of the transcripts from the lichen samples. Raw reads from both libraries were deposited in the NCBI database with the accession number SRP138994.
Penggunaan enzim selulase untuk penguraian biojisim pertanian lignoselulosa telah lama dikaji dan pelbagai usaha telah dilakukan untuk meningkatkan kecekapan proses hidrolisis. Keberkesanan penguraian biojisim pertanian kepada gula ringkas memerlukan satu campuran enzim yang mengandungi pelbagai jenis aktiviti selulolitik. Dalam kajian ini, satu campuran multi-enzim rekombinan yang terdiri daripada tiga komponen asas selulase iaitu endoglukanase (EglB) dan β-glukosidase (BglA) daripada Aspergillus niger serta selobiohidrolase (CbhII) daripada Trichoderma virens telah dibentuk khusus untuk hidrolisis tandan kosong kelapa sawit (TKKS). Penghasilan enzim selulase rekombinan telah dilakukan menggunakan hos pengekspresan Pichia pastoris. Pengoptimuman nisbah enzim untuk tindak balas ditentukan menggunakan Kaedah Gerak Balas Permukaan (RSM). Hasil menunjukkan hidrolisis TKKS pada suhu 50 °C dan pH 5.0 menggunakan enzim pada nisbah 641.4 unit CMCase: 10.14 unit Avicelase: 93.8 unit β-glukosidase, menghasilkan gula terturun dan glukosa tertinggi, masing-masing sebanyak 63 mg dan 40 mg per gram substrat TKKS. Hasil hidrolisis TKKS oleh campuran multi enzim yang telah dibentuk dalam kajian ini menunjukkan ketiga-tiga gabungan enzim rekombinan ini berpotensi untuk digunakan bagi penguraian TKKS.