Aims:The objective of this research was to isolate caffeine-degrading bacteria from coffee pulp waste in Indonesia and characterize their caffeine degradation activity. Methodology and results: The caffeine-degrading bacteria were isolated from coffee pulp wastes of Coffea arabica and C. canephora. These isolates were selected based on their caffeine degradation activity. The identification and biochemical properties of the best isolate were conducted via 16S rDNA sequence analyses and by using the Microbact kit. Meanwhile, caffeine degradation activity of this bacteria was analyzed by using LC-MS/MS. The results indicated that fourteen bacterial isolates were able to degrade caffeine. The highest caffeine degradation activity was performe d by isolate KRM9 at the rate of 99.26 ± 0.01%, on a caffeine medium after 24 h of incubation. Based on the 16S rDNA analyses, the KRM9 isolate was identified as Pseudomonas monteilii. Till present, this species has not been reported as a caffeine-degrading bacterium. However, LC-MS/MS analysis indicated that caffeine was degraded by P. monteilii KRM9 and theobromine was not the secondary metabolite of caffeine degradation. Conclusion, significance and impact of study: Pseudomonas monteilii KRM9 was detected as a new isolate of caffeine-degrading bacteria. This bacterium can be introduced as an agent to degrade caffeine from coffee pulp waste. It is expected that further research can be conducted on the overall mechanism of caffeine degradation by P. monteilii KRM9.

Aims:The objective of the research was to get the potential cellulolytic bacteria which was caffeine tolerance from Indonesian coffee pulp waste. Methodology and results: The cellulolytic bacteria were isolated from coffee pulp wastes of Coffea arabica and C. canephora. These isolates were selected based on their cellulose hydrolysis, CMCase activity, and caffeine tolerance. The density of cellulolytic bacteria of C. arabica pulp waste was 4.7 ± 3.5 × 10 6 CFU/g, and that of C. canephora pulp waste was 1.5 ± 1.5 × 10 6 CFU/g. Among 61 cellulolytic bacterial isolates, 24 isolates formed clear zones on CMC medium with Gram iodine flooding. Three isolates (CRM10, CRM1, and CRM12) from C. canephora pulp waste had the highest cellulolytic activity. Based on the CMCase activity, it was indicated that an isolate of CRM10 showed the highest CMCase activity with 3.38 ± 0.65 U/mL. This bacteria had tolerance ability to caffeine until 0.4% on nutrient agar medium. Isolates of CRM10 had similarity to Bacillus subtilis based on 16S rDNA sequence. Conclusion, significance, and impact of study: CRM10 was identified as Bacillus subtilis and considered as a potential isolate to degrade cellulose of coffee pulp waste that contained caffeine. .

Bacteria are the most dominant group of microorganisms in aquatic environments due to their role in organic matter decomposition. Decomposition activity is related to the type and dominance of bacteria in the communities. Therefore, study of bacterial diversity is an important step to understand their role in aquatic ecosystems. This study was to determine bacterial diversity and their physiological characters of bacteria from Bandealit Coast in Jember East Java Indonesia. The bacteria were confirmed by BOX-PCR profile for their genetic polymorphisms. Identification of potential isolate was conducted based on 16S rRNA gene sequence. The result showed that BA011109 isolate was able to utilize D-cellobiose as a sole substrate, indicating its ability to hydrolyse β β β β βglucoside bond. This isolate was a potential decomposer in the area considering that most of organic pollutants were from plants that cointain high cellulose. Based on its 16S rRNA gene sequence, this isolate was closely related to Microbacterium esteraromaticum with 100% homology. Further study on quantitative hydrolytic activities is needed to elucidate its role as an organic matter decomposer in aquatic environment.

A blood clot (thrombus) in a blood stream is formed due to a circulatory system imbalance in the hemostasis which results in plug of blood vessels. The suppliy of nutrients and oxygen to the tissues is inhibited (ischemia) by the accumulation of thrombus and embolus in the blood vessel. This prosses is the main cause for further atherotrombotic diseases such as myocardial infraction and cerebral infraction. This disease could be overcome by thrombolytic therapy by using fibrinolytic protease enzyme. Fibrinolytic activity of protease enzymes have been studied from various species of bacteria. Bacterial isolate of WU 021055* obtained from Papuma coastal waters has demonstrated fibrinolytic activity. This research was aimed to identify the bacterial isolate through morphological characterization (colony and cell morphology), physiological characterization (indole test, carbohydrates fermentation test (glucose, lactose, sucrose and fructose), catalase test, starch hydrolysis test, and the pH effect test), and molecular identification using 16S rRNA. Based on those characterizations, the bacterial isolate of WU 021055* shows a high similarity to Bacillus aerius.Keywords: Atherotrombosis, fibrinolytic, identification, characterization, bacteria ABSTRAKBekuan darah (trombus) dalam peredaran darah terbentuk akibat ketidakseimbangan sistem sirkulasi dalam hemostasis yang menyebabkan penyumbatan pembuluh darah. Akumulasi trombus dan embolus pada pembuluh darah mengakibatkan suplai nutrisi dan oksigen ke jaringan terhambat (iskemia) dan bahkan kematian jaringan (infark). Pembentukan ini merupakan etiologi dari penyakit aterotrombosis seperti infark miokard dan infark serebral. Penyakit akibat trombosis ini dapat diatasi dengan terapi trombolitik dengan enzim protease fibrinolitik. Aktivitas enzim protease fibrinolitik telah diteliti dari berbagai spesies bakteri. Isolat bakteri WU 021055* asal perairan pantai papuma tampak memiliki aktivitas fibrinolitik. Pada penelitian ini dilakukan identifikasi isolat bakteri melalui karakterisasi morfologi (morfologi koloni dan sel), karakterisasi fisiologis (uji indol, uji fermentasi karbohidrat (glukosa, laktosa, sukrosa dan fruktosa), uji katalase, uji hidrolisis pati, dan uji pengaruh pH), dan identifikasi secara molekuler menggunakan 16S rRNA. Berdasarkan karakterisasi morfologi, fisiologi, dan marker 16S rRNA, isolat bakteri WU 021055* menunjukkan kemiripan yang tinggi dengan Bacillus aerius.Kata kunci: Aterotrombosis, fibrinolitik, identifikasi, karakterisasi, bakteri

Naturally, the coffee pulp contains caffeine. Caffeine-degrading bacteria can use caffeine for their growth. The purpose of this research was to obtain high potency of caffeine-degrading bacterial isolates. These bacteria were isolated from naturally-fermented pulp waste of Coffea arabica. The bacteria were isolated by using minimal medium M9 contain 1 g/L caffeine and was screened based on their activity to degrade caffeine. There 13 isolates that successfully isolated by this medium. The research found five isolates had high potency to degrade caffeine. They were KAFS 33, KAFS 34, KAFS 16, KAFS 47, and KAFS 35 respectively. Those isolates were the potential to be further analysis as an agent of decaffeinating coffee.