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ABSTRACT Wine coffee is fermented coffee cherries with a wine-like flavor and aroma. The purpose of this study was to isolate, characterize ethanol-tolerant and thermotolerant yeast from Semeru Arabica wine coffee cherries and to determine the molecular identification of the selected yeast. In this study, the Semeru Arabica coffee cherries were fermented for 30 days.Yeast isolation was performed on samples collected each week during fermentation. The selected isolates were characterized for macroscopic and microscopic morphology, followed by ethanol and temperature tolerance analysis. The isolate with the most desirable characteristics, designated CW30, grew well at 30ºC but was sensitive to temperatures above 35ºC. The isolate grew well in media containing 5% ethanol but was sensitive to a 10% ethanol concentration. Based on nucleotide sequence analysis, CW30 exhibited 100% similarity with Candida ethanolica isolate 3-1-19. Keywords: Fermented coffee, Semeru Arabica, Wine coffee, Yeast isolation

ABSTRACT This study was conducted to identify the best fermentation period and roasting temperature for semeru arabica wine coffee. This study used the factorial RSBD research approach (three levels of roasting temperature and four levels of fermentation duration). Physicochemical and organoleptic data were analyzed using ANOVA 5%, LSD 5%, and HSD 5%. Then, the best treatment's ash alkalinity, coffee juice content, and caffeine level were assessed. According to the study, the fermentation period has impact on the moisture level of green beans, the moisture and ash content of roasted beans, acidity, body, and balance. The water and ash content of roasted beans, aroma, flavor, acidity, aftertaste, body, balance, and overall attribute, are influenced by the roasting temperature. Then the interaction between factors significantly affects the water content of roasted beans, flavor, acidity, and body. The best treatment achieved a total score of 81.83 (40 days, 200 °C). Keywords: Coffee characteristics, Fermentation, Roasting, Wine coffee

ABSTRACT The current trend in coffee product diversification is wine coffee, which has a wine-like flavor and aroma. Reviews of wine coffee are still limited, especially those related to microbial succession during the fermentation process. The objective of this research was analyzing the microbial population and the effect of microbial succession during fermentation process of Arabica Semeru coffee. The analysis was conducted by identifying microbes microscopically and macroscopically. The results of microbial identification during the 30-day fermentation showed populations of yeast, lactic acid bacteria, and acetic acid bacteria. Along with the length of fermentation time, the temperature decreased and the pH fluctuated and stabilized after 16 days of fermentation. There was an increase in sugar and protein content, from 0.61% to 2.87% and 2.13% to 2.23%, respectively. The ethanol content reached 0.047% at the end of fermentation. The microbial exploration was required for further investigation in understanding its role during fermentation process. Keywords: Arabica Semeru, Bacteria, Fermentation, Succession, Wine coffee, Yeast

Polyethylene terephthalate (PET) plastic is the most widely used type of plastic that produces waste and causes various environmental and health problems. The treatment of PET plastic waste with chemically and mechanically recycling approaches still has shortcomings, so biological processing using microorganisms or enzymes has new potential. Two bacterial isolates from the Indonesian Culture Collection of National Research and Innovation Agency (InaCC, BRIN), namely isolate InaCC B538 and InaCC B947, were further observed for their potential in PET plastic degradation. Firstly, both isolates were determined by the molecular marker 16S rDNA. The potential of both isolates was measured with following method: 10 days of degradation using PET and PCL substrates, esterase enzyme activity assay, and observation of the PET plastic surface using Scanning Electron Microscope (SEM). Species identification was performed using DNA sequencing of 16S rDNA. InaCC B538 and InaCC B947 were closely related to Bacillus altitudinis TBMAX41 and Alcaligenes faecalis AN-13, respectively. InaCC B947 isolate has a better potential in degrading PET plastic and PCL with a degradation percentage of 0.32% for PET plastic and 3.22% for PCL film for 10 days, respectively, and esterase activity of 0.06 U/ml; while InaCC B538 did not cause weight loss of PET and 2.49% for PCL, respectively, with esterase activity of 0.04 U/ml. The degradation of PET plastic by the isolates InaCC B947 was able to cause damage to the plastic surface leading to the degradation of PET plastic.

Photoaging is skin aging caused by exposure to UV rays, which increases the expression of matrix metalloproteinase-1 (MMP-1). The photoaging process is related to the degradation of collagen types I and III in the extracellular matrix by MMP-1, which causes wrinkles on the skin. MMP-1 inhibitors from natural products have the potency as anti-photoaging. This study aims to screen the potency of bioactive compounds from Impatiens balsamina L. as MMP-1 inhibitors through in silico studies. The best test ligands were selected based on bioavailability, pharmacokinetics, toxicity, and molecular docking tests against the target protein MMP-1 (PDB ID: 1HFC) compared to that of control ligands (PLH and doxycycline). Peonidin, kaempferol, and pelargonidin were selected as the best test ligands because they accomplish the characteristics of bioavailability, pharmacokinetics, and toxicity. Based on molecular docking results, those test ligands have better binding affinity than that of control ligands, as indicated by rerank scores of -108.807 kcal/mol, -99.9796 kcal/mol, and -98.9128 kcal/mol, respectively. Those test ligands also formed the same interactions with control ligands at residues Ala182, Asn180, Glu219, and Leu181. The results suggest peonidin, kaempferol, and pelargonidin were candidates for anti-photoaging agents through MMP-1 inhibition.

To overcome these problems, our team, initiate the establishment of SMEs involving students at the Babussalam Al-Firdaus Islamic Boarding School through diversification of dairy-based processed products and bakery products. Introduction of some lectures including bakery products technology, milk processing technology, packaging and storage technology, food quality assurance and Good Manufacturing Practice (GMP), product trials and training activities are carried out offline at the Food Production and Training Center (FPTC), Universitas Brawijaya. The new SMEs empowerment of students and provide additional income for the Babussalam Al-Firdaus Islamic Boarding School to be financial independence.

Abstract Background 2,3-Butanediol (2,3-BD), a valuable compound used for chemicals, cosmetics, pesticides and pharmaceuticals, has been produced by various microbes. However, no high-temperature fermentation of the compound at high productivity has been reported. Methods Thermotolerant xylose-utilizing microbes were isolated from 6 different districts in Laos and screened for a low accumulation of xylitol in a xylose medium at 37 ˚C. One isolate was found to produce 2,3-BD and identified by 16S rDNA sequencing. The 2,3-BD fermentation capacity was investigated at different temperatures using xylose and glucose as carbon sources, and the fermentation parameters were determined by a high-performance liquid chromatography system. Results By screening for a low accumulation of xylitol in a xylose medium, one isolate that accumulated almost no xylitol was obtained. Further analyses revealed that the isolate is Cronobacter sakazakii and that it has the ability to produce 2,3-BD at high temperatures. When xylose and glucose were used, this strain, named C. sakazakii OX-25, accumulated 2,3-BD in a short period before the complete consumption of these sugars and then appeared to convert 2,3-BD to acetoin. The optimum temperature of the 2,3-BD fermentation was 42 ˚C to 45 ˚C, and the maximum yield of 2,3-BD was 0.3 g/g at 12 h in 20 g/l xylose medium and 0.4 g/g at 6 h in 20 g/l glucose medium at 42 ˚C. The 2,3-BD productivity of the strain was higher than the 2,3-BD productivities of other non-genetically engineered microorganisms reported previously, and the highest productivity was 0.6 g/l·h and 1.2 g/l·h for xylose and glucose, respectively. Conclusions Among thermotolerant microbes isolated in Laos, we discovered a strain, C. sakazakii OX-25, that can convert xylose and glucose to 2,3-BD with high efficiency and high productivity at high temperatures, suggesting that C. sakazakii OX-25 has the potential for industrial application to produce 2,3-BD as an important platform chemical.

Currently, Cardiovascular Diseases (CVDs) cases are increasing significantly in Indonesia and the world. WHO reported that in 2019-related CVD patients accounted for 85%, which increased to 87% in 2021. The cases increased significantly by 2%, which is a significant number as it covered the world population. Stroke is a disease affected by platelets undergoing thrombolysis by the α-thrombin (IIa) receptor with coagulation factors on the heart blood vessel. Chemical or synthesis drugs to treat thrombolytics such as heparin are already available, but they would be harmful if misused. In this study, we focused on the activity, interaction, and prediction of the physicochemical of anti-thrombotic peptides. We found that k-casein from kefir has the highest activity of 0.037 A based on BIOPEP analysis with 17 peptide libraries and molecular docking visualization analysis using PyMol, PyRx, and Discovery Studio for screening the best peptide. ToxinPred and AlgPred showed there are no potentially harmful peptides, but the protein variant of β-conglycinin subunit can be an allergen known to the immune system IgE. In conclusion, the GPR peptide has the highest binding affinity, with a total energy of -6.8 kcal/mol.

Cocoa bean shell is a by-product of processing chocolate products that can be used as a tea infusion because it contains bioactive compounds such as polyphenols which act as antioxidants. Roasting plays a very important role in the development of aromas and flavors. The temperature commonly used in roasting is about 100 – 150 ºC. Phenolic content plays a role in the formation of sensory properties of the product due to the presence of interaction with proteins through the Maillard reaction that occurs in the roasting process. This study aims to determine the effect of roasting temperature on the antioxidant activity of tea infusions from cocoa bean shells. The methods used in this study are the folin-ciocalteu method and the DPPH method. Cocoa bean shell tea with a roasting temperature of 100ºC, 20 minutes has the highest total phenols, antioxidant levels, and radical scavenging activity of 21.54 ± 0.37 mg GAE/g; 12.80 ± 0.17 mg AA/g; and 85.18 ± 1.28%. The total amount of phenols has a very strong correlation to antioxidant levels, radical scavenging, the color value of the brew L*, b*, and is inversely proportional to the color value of a*. Cocoa bean shell tea roasting temperature of 140ºC, 18 minutes has the highest IC50 value of 22.76 mg /ml. In addition, the roasting temperature can affect the sensory attribute character of cocoa bean shell tea.

Penelitian ini bertujuan memperoleh isolat khamir penghasil etanol yang memiliki toleransi tinggi terhadap etanol dan suhu. Tahapan dalam penelitian ini meliputi isolasi khamir, identifikasi morfologi koloni dan sel, analisis kurva pertumbuhan khamir, skrining khamir thermotolerant dan ethanol-tolerant, fermentasi dan analisis kadar etanol. Isolat JCBI-01 (Kenitu) dan JCBI-14 (Coklat) diduga genus Candida sp. Sedangkan isolat JCBI-02 (Pisang) dan JCBI-03 (Pepaya) diduga genus Saccharomyces sp. Pada uji ethanol-tolerant isolat JCBI-01, JCBI-02, JCBI-14, dan JCBI-03 memiliki toleransi yang baik terhadap etanol konsentrasi 14%, dan pada isolat JCBI-01 dan JCBI-14 masih dapat toleran pada konsentrasi 15%. Hasil uji thermotolerant semua isolat bertahan hingga suhu 48 ̊C. Untuk kemampuan dalam produksi etanol pada jam ke-48, isolat JCBI-01 menghasilkan etanol sebesar 2.41% (b/v),JCBI-02 sebesar 2.54% (b/v), JCBI-14 sebesar 2.63% (b/v), dan JCBI-03 sebesar 3.07% (b/v).

Abstract Kmmig1 as a disrupted mutant of MIG1 encoding a regulator for glucose repression in Kluyveromyces marxianus exhibits a histidine-auxotrophic phenotype. Genome-wide expression analysis revealed that only HIS4 in seven HIS genes for histidine biosynthesis was down-regulated in Kmmig1. Consistently, introduction of HIS4 into Kmmig1 suppressed the requirement of histidine. Considering the fact that His4 catalyzes four of ten steps in histidine biosynthesis, K. marxianus has evolved a novel and effective regulation mechanism via Mig1 for the control of histidine biosynthesis. Moreover, RNA-Seq analysis revealed that there were more than 1,000 differentially expressed genes in Kmmig1, suggesting that Mig1 is directly or indirectly involved in the regulation of their expression as a global regulator.

Candida tropicalis, a xylose-fermenting yeast, has the potential for converting cellulosic biomass to ethanol. Thermotolerant C. tropicalis X-17, which was isolated in Laos, was subjected to repetitive long-term cultivation with a gradual increase in temperature (RLCGT) in the presence of a high concentration of glucose, which exposed cells to various stresses in addition to the high concentration of glucose and high temperatures. The resultant adapted strain demonstrated increased tolerance to ethanol, furfural and hydroxymethylfurfural at high temperatures and displayed improvement in fermentation ability at high glucose concentrations and xylose-fermenting ability. Transcriptome analysis revealed the up-regulation of a gene for a glucose transporter of the major facilitator superfamily and genes for stress response and cell wall proteins. Additionally, hydropathy analysis revealed that three genes for putative membrane proteins with multiple membrane-spanning segments were also up-regulated. From these findings, it can be inferred that the up-regulation of genes, including the gene for a glucose transporter, is responsible for the phenotype of the adaptive strain. This study revealed part of the mechanisms of fermentability at high glucose concentrations in C. tropicalis and the results of this study suggest that RLCGT is an effective procedure for improving multistress tolerance.

Tujuan dari penelitian ini adalah mengisolasi dan mengidentifikasi mikroorganisme penghasil enzim xilanase dari lumpur panas Lapindo. Selain itu penelitian ini bertujuan untuk mengetahui karakteristik enzim xilanase yang dihasilkan. Isolat yang diperoleh dimungkinkan merupakan mikroorganisme termofilik berdasarkan kondisi suhu tinggi di lumpur panas. Penelitian ini terdiri dari pengambilan sampel, isolasi, karakterisasi isolat termofilik dan karakterisasi enzim xilanase yang dihasilkan. Sebanyak 14 isolat berhasil didapatkan dan isolat C211 menunjukkan aktivitas hidrolisis terbaik. Hasil dari identifikasi molekuler berbasis 16S rRNA menunjukkan memiliki kedekatan genetik dengan B. lichenformis dan B. aerius dengan derajat kesamaan homolog masing-masing 99% dan 98%, mengindikasikan bahwa isolat tersebut termasuk genus Bacillus. Isolat C211 merupakan tipe Gram positif dan menunjukkan hasil positif pada uji Voges Proskauer, indol, dan ornithine. Suhu pertumbuhan berada pada suhu 45-65ºC dengan kondisi pH 5.8-7.2. Enzim xilanase kasar dari isolat C211 memiliki aktivitas optimum pada suhu 50ºC dan pH 7 sebesar 3.95 Unit/mL.

Arabinofuranosidase (Abfa) is one of the most important enzymes involved in degradation of lignocelullose biomass. Two genes encoding α-L-Arabinofuranosidase (abfA), each from Bacillus subtilis DB104 (abfAa1) and an indigenous Indonesian B. licheniformis CW1 (abfAb3), were cloned by the PCR approach and expressed in Escherichia coli. Sequences analysis of abfAa1 and abfAb3 revealed that each consists of 1721 and 1739 base pairs long DNA, respectively. Each clone contains a hypothetical open reading frame of 1503 and 1509 bp that encode an Abfa protein of 500 and 502 amino acids for abfAa1 and abfAb3, respectively. The deduced amino acid sequence of AbfaB3 shares 75% identity to that of AbfaA1. The recombinant enzymes were expressed constitutively in E. coli. Partial characterization of those enzymes revealed that the AbfaA1 and AbfaB3 were optimally active at 50 ºC and 60 ºC at pH 6, respectively. Thermostability studies of the recombinant enzymes with p-nitrophenyl α-L-arabinofuranoside at their optimal conditions showed that up to 50% AbfaA1 activity was lost after 5 h incubation at 50 ºC, whereas the AbfaB3 retained its activity over 75% after 12 h pre-incubation oat 60 ºC. This thermostability study of recombinant AbfaB3 showed for the first time that the arabinofuranosidase from B. licheniformis is a thermostable enzyme. The recombinant enzyme showed a higher optimal reaction temperature (60 ºC) in comparison to the previously reported thermostable arabinofuranosidase. The thermostable AbfaB3 has a potential to be applied to the degradation of lignocellulose biomass synergistically with thermostable xylanases, for instance in the production of xylo-oligosaccharides.