Macroalgae and macroalgae-associated bacteria together constitute the most efficient metabolic cycling system in the ocean. Their interactions, especially the responses of macroalgae-associated ...bacteria communities to algae in different geographical locations, are mostly unknown. In this study, metagenomics was used to analyze the microbial diversity and associated algal-polysaccharide-degrading enzymes on the surface of red algae among three remote regions. There were significant differences in the macroalgae-associated bacteria community composition and diversity among the different regions. At the phylum level, Proteobacteria, Bacteroidetes, and Actinobacteria had a significantly high relative abundance among the regions. From the perspective of species diversity, samples from China had the highest macroalgae-associated bacteria diversity, followed by those from Antarctica and Indonesia. In addition, in the functional prediction of the bacterial community, genes associated with amino acid metabolism, carbohydrate metabolism, energy metabolism, metabolism of cofactors and vitamins, and membrane transport had a high relative abundance. Canonical correspondence analysis and redundancy analysis of environmental factors showed that, without considering algae species and composition, pH and temperature were the main environmental factors affecting bacterial community structure. Furthermore, there were significant differences in algal-polysaccharide-degrading enzymes among the regions. Samples from China and Antarctica had high abundances of algal-polysaccharide-degrading enzymes, while those from Indonesia had extremely low abundances. The environmental differences between these three regions may impose a strong geographic differentiation regarding the biodiversity of algal microbiomes and their expressed enzyme genes. This work expands our knowledge of algal microbial ecology, and contributes to an in-depth study of their metabolic characteristics, ecological functions, and applications.
Cardiovascular disease is among the largest contributors of premature mortality in the world caused by inflammation of blood vessels. The abnormalities provoke thrombus formation or thrombosis ...blocking blood vessels leading to strokes, heart attacks and coronary artery diseases. Increasing percentage of cardiovascular cases and deaths due to thrombosis has attracted researchers to look for newer thrombolysis agents. Commonly used drugs to treat thrombosis has been limited due to various side effects. Therefore, the search for sources of safer and cheaper fibrinolytic enzymes for handling thrombolysis continues. This study aimed to evaluate potentials of fibrinolytic protease of bacteria isolated from fermented seafood (sea cucumber) products as antithrombotic agents. Information was initially gathered from scientific publications identified using web-based tools including PubMed (National Center for Biotechnology Information), Science Direct (Scopus) and Web of Science (Thomson Reuters) using combinations of search terms including “fibrinolytic enzyme protease”, “endopeptidase”, “fermented food”, “sea cucumber”, “thrombolysis therapy,” “thrombolytic agent,” “fibrinolytic bacteria,” “fibrinolysis,” “protease producing bacteria,” “fibrin degradation,” “holothurians,” etc. We also searched for these terms in national and international organization technical reports and databases. This literature review reveals the prospects of fibrinolytic protease enzymes from bacteria from fermented seafood, particularly sea cucumber as novel antithrombotic agents.
An extracellular agarase was purified from Bacillus sp. BI-3, a thermophilic agar-degrading bacterium isolated from a hot spring in Indonesia. The purified agarase revealed a single band on sodium ...dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular mass of 58 kDa. The optimum pH and temperature of the agarase were 6.4 and 70°C, respectively. The activity of the agarase was stable at high temperatures, and more than 50% activity was retained at 80°C for 15 min. Furthermore, the enzyme was stable in the pH range of 5.8-8.0, and more than 60% of the residual activity was retained. Significant activation of the agarase was observed in the presence of K(+), Na(+), Ca(2+), Mg(2+), and Sr(2+); on the other hand, Ba(2+), Zn(2+), Cu(2+), Mn(2+), Co(2+), Fe(2+), and EDTA inhibited or inactivated the enzyme activity. The components of the hydrolytic product analyzed by thin-layer chromatography showed that the agarase mainly produced neoagarobiose. This study is the first to present evidence of agarolytic activity in aerobic thermophilic bacteria.
Glucose oxidase was an enzyme which catalyzed β-D-Glucose to gluconic acid and hydrogen peroxide. Glucose oxidase from Aspergillus niger IPBCC.08.610 was isolated, purified and characterized. The ...enzyme was purified by ammonium sulphate precipitation and dialysis. The specific activity and yield of dialysis fraction were 19.766 U/mg and 4.744%. The optimum pH and temperature were 6 and 30oC. The stability of enzyme at optimum pH and temperature was decreasing 50% at 25 minutes. The km and vmax values for enzyme were 27 mM and 0.986 U/mg.
Karakteristik hidrolisat protein ikan (HPI) dipengaruhi oleh kondisi proses hidrolisisnya. Optimasi produksi HPI kuniran (Upeneus sulphureus) telah dilakukan secara enzimatis dengan perlakuan ...perbandingan ikan dan air (1:1 dan 1:2) dan penambahan enzim protease 500 U, 1.000 U, dan 1.500 U per 25 g ikan. Nilai derajat hidrolisis (DH) HPI digunakan untuk menentukan kondisi optimum produksi HPI. Berdasarkan kondisi optimum tersebut, produksi HPI skala diperbesar dilakukan dengan 500 g ikan kuniran sebagai bahan baku. Karakteristik HPI skala diperbesar yang diamati adalah karakteristik kimia (air, abu, protein, lemak, asam amino, daya cerna protein, peptida, dan berat molekul) dan fisik (warna, aktivitas, dan stabilitas emulsi). Hasil penelitian menunjukkan bahwa perbandingan ikan, air, dan jumlah enzim berpengaruh terhadap nilai DH HPI. Pada skala produksi HPI diperbesar, karakteristik kimia dari HPI kuniran secara umum dipengaruhi oleh perbedaan perlakuan hidrolisis, tetapi tidak sifat fisiknya. Perbedaan kandungan protein, abu, dan peptida dari HPI lebih dominan dipengaruhi oleh perbedaan perlakuan perbandingan ikan dan air, sedangkan kombinasi perlakuan jumlah enzim dan perbandingan ikan dan air berpengaruh terhadap kandungan asam amino dan daya cerna protein HPI. Secara umum, perlakuan hidrolisis menggunakan enzim 1.000 U/25 g dan perbandingan ikan dan air 1:1 (kode E1000A11) memberikan perlakuan terbaik yang terlihat dari kandungan protein, asam amino, peptida, dan daya cerna tertinggi dibandingkan perlakuan lain. Kandungan kimia dan daya cerna yang tinggi tersebut memberikan peluang aplikasi HPI sebagai penambah nutrisi bagi balita. ABSTRACT The characteristics of fish protein hydrolyzate (FPH) are affected by their hydrolysis process conditions. Production optimization of FPH kuniran (Upeneus sulphureus) has been carried out enzymatically with treatments of fish and water ratio (1: 1 and 1: 2) and protease enzymes 500 U; 1,000 U, and 1,500 U per 25 g fish. The degree of hydrolysis (DH) value of FPH was used to determine the optimum conditions of FPH production. Based on this optimum condition, the enlarged production of FPH (500 g of fish) was carried out. Chemical characteristics (moisture, ash, protein, fat, amino acids, protein digestibility, peptide, and molecular weight) and physical (color, the emulsion activity, and stability) of FPH from enlarged production were analyzed. The results showed that the ratio of fish and water and the amount of enzyme affect the DH value of FPH. At the enlarged scale, the chemical characteristics of FPH kuniran were generally influenced by different hydrolysis treatments but not their physical properties. The differences in protein, ash, and peptide content of FPH were more dominantly affected by differences in the fish and water ratio, while the combination treatments of the enzymes and the ratio of fish-water affected the amino acid content and protein digestibility of FPH. In general, the hydrolysis treatment using a 1,000 U enzyme/25 g fish and the ratio of fish and water 1:1 (code E1000A11) was the optimum treatment as seen from the highest protein, amino acid, peptide, and protein digestibility compared to other treatments. The high chemical content and digestibility of FPH provide an opportunity application of FPH as a nutritional enhancer for toddlers.
There is no report of diversity, biological properties and bioactive compounds of sponge-associated fungi from Indonesia’s mangrove ecosystem to date. This study was designed to isolate ...sponge-associated fungi from a mangrove ecosystem in Mangkang, to screen the antimicrobial and extracellular enzyme properties of the isolates, characterize the biologically promising isolates using molecular approaches, and profile the secondary metabolites using phytochemical and thin-layer chromatography (TLC) analyses. An unidentified sponge that lived in association with mangrove roots was collected from Mangkang. Total of eight associated fungi were isolated from the sponge. Among all isolates, only two fungi SPMKF 1 and SPMKF 6 produced extracellular amylase, another two fungi SPMKF 4 and 5 showed antibacterial activity against MRSA, and only one fungus SPMKF 8 was able to produce extracellular amylase and show antimicrobial activity against ESBL
E
.
coli
,
Salmonella enterica
ser. Typhi strain MDR and
C
.
albicans
, while SPMKF 2, SPMKF 3 and SPMKF 9 did not show any biological properties. The result of genetic characterization proved that SPMKF 1 was
Cladosporium tenuissimum
, SPMKF 4 was
Eutypella
sp., SPMKF 5 was
Lasiodiplodia theobromae
, SPMKF 6 was
Fusarium keratoplasticum
and SPMKF 8 was
F
.
solani
. Furthermore, an amylase gene was detected in fungi SPMKF 1, 6 and 8 while among the BGC, only NRPS genes were detected in SPMKF 4, 5 and 8. Interestingly, several same metabolites indicating the same retention factor (
R
f
) values in TLC were detected in the fungal crude extracts by TLC.
Cardiovascular disease is the primary cause of mortality in the world due to the formation of blood clots or thrombi in blood vessels. Bacterial proteases commonly function as thrombus dissolver ...agents in the pharmaceutical industry. Bacterial isolate HSFI-10 (Holothuria scabra Fermented Intestine-10) previously isolated from Rusip fermented sea cucumber had demonstrated thrombolytic activity. This study aimed to produce crude protease of HSFI-10 strain at an optimized incubation time and determine the thrombolytic activity of crude and dialysate proteases on A, B, AB, and O blood types. Isolate HSFI-10 was first molecularly identified and found to be Bacillus cereus with a homology level of 99.80% with Bacillus cereus strain ST06. The optimum crude enzyme was obtained after 48-h incubation with an activity of 222.52 U/mL, which increased to 438.84 U/mL after ammonium sulfate precipitation and dialysis. Clot lysis activity of crude enzymes was measured based on the gravimetry method on blood in the ABO system, showing results that ranged from 68.99% to 69.76%, while the dialysate ranged from 81.16% to 82.52%. In conclusion, partial purification of bacterial protease could increase both its specific and thrombolytic activities on human blood in the ABO system, with only 1% activity variability between A, B, AB, and O blood types.
Seaweed is a rich source of phycocolloids, which include agar, alginate, and carrageenan. Low molecular weight polysaccharides, namely oligomers or oligosaccharides, can be produced from seaweed ...polysaccharides through enzymatic degradation. Most of these enzymes are produced by microorganisms closely associated with seaweed. This study aimed to isolate and select polysaccharidases-producing (SPases) bacteria associated with wild seaweed from the sea around Ambon Island, an area famous for its high marine biodiversity index. A total of 11 types of marine algae samples could be collected, and as many as 92 bacterial isolates could be cultured from all these algae samples. Screening used a clear zone method on a solid medium containing substrates agar, alginate, or carrageenan and followed by Lugol’s iodine staining solution showed that a total of 74 of the 92 bacterial isolates obtained were SPases-producing with the composition: agarase-producing (28 isolates), and alginate lyase-producing (26 isolates), and carrageenaseproducing (20 isolates). The 16S rRNA identification results showed that the 74 bacterial isolates were representative of 13 species and belong to 2 classes, namely Gammaproteobacteria and Bacillus. The bacterial isolates in the Gammaproteobacteria class obtained consisted of three genera: Pseudoalteromonas (32 isolates), Cobetia (18 isolates), and Microbulbifer (15 isolates). Bacterial isolates in the Bacillus class obtained only contain a genus consisting of 8 isolates. In conclusion, the sea around Ambon Island is a potential source of polysaccharidases-producing algal symbiont bacteria.
Abstract
Among essential treatment of cardiovascular disorders are fibrinolytic proteases. Most thrombolysis agents are fibrinolytic enzymes from group of bacterial proteases. This work reports a ...potent bacterium isolated from fermented intestine of
H. scabra
, which could produce fibrinolytic protease with high thrombolysis activity. Bacterial selection was conducted based on proteolytic and fibrinolytic activities indicated as clear zone on skim milk and fibrin agar media, respectively. Crude proteases from the selected bacterial isolates were subjected to thrombolytic activity test based on gravimetric method, which results were confirmed after 7 repetitions. As result, 4 fibrinolytic protease-producing bacterial isolates HFSI-3, HFSI-4, HFSI-5 and HFSI-8 were obtained. Among them, HFSI-5 isolate identified as
Bacillus tequilensis
on the basis of the 16S rRNA gene sequencing and morphological properties produced crude protease with the highest thrombolytic activity. The thrombolytic activity of crude protease produced by
B. tequilensis
HFSI-5 is worthy of comparing to that of standard fibrinolytic enzyme Nattokinase showing its potential as thrombolysis agent.
Background
Marine bacteria have recently attracted increasing attention to be harnessed for the production of valuable enzymes, vitamins, and bioactive compounds. Bacteria associated with the ...surfaces of marine macroalgae, called epibionts, are particularly interesting from ecological and biotechnological points of view, as they often exhibit antimicrobial activities to compete with pathogenic bacteria for nutrients and spaces. In search for biotechnologically potential genes from marine bacteria, we sequenced and analysed the genome of the epibiont HI03-3b, a polysaccharide-degrading bacterium associated with the surface of the Indonesian brown algae
Hydroclathrus
sp.
Results
The algal epibiont HI03-3b has a genome of approximately 4,860,704 bp in size with 42.02 mol% G + C content, consisting of 5655 open reading frames (ORFs), 4409 genes coding for proteins (CDSs), 94 genes for tRNAs, and 32 genes for rRNAs. The genome sequence of HI03-3b was most closely related to that of
Cytobacillus firmus
NCTC10335 with the average amino acid identity (AAI) of 95.0 %, average nucleotide identity (ANI) of 94.1 %, and a recommended DNA-DNA hybridization (DDH) of 57.60 %. These scores are lower than the most frequently used standard for species demarcation (95% ANI cutoff) and the new species threshold (DDH > 70.0% for the same bacterial species). Some differences in genome features and gene composition were observed between HI03-3b and NCTC10335, such as genes encoding carbohydrate active enzymes. These suggest that HI03-3b is unique and likely a novel species within
Cytobacillus
genus, and we therefore proposed its name as
Cytobacillus wakatobiense
HI03-3b. Genome sequence analyses indicated the presence of genes involved not only in polysaccharide and protein degradation but also in vitamin and secondary metabolite biosynthesis. Some of them encode enzymes and compounds with biotechnological interest, such as protease, chitinase, subtilisin, pullulanase, and bacillolysin, which are often associated with antimicrobial or antibiofilm activities. This antimicrobial potential is supported by our finding that the extracellular protein fraction of this epibiont inhibited the growth of the bacterial pathogen
Staphylococcus aureus
.
Conclusion
The epibiont
Cytobacillus
HI03-3b harbours genes for polysaccharide and protein degradation as well as for natural product biosynthesis, suggesting its potential ecological roles in outcompeting other bacteria during biofilm formation as well as in protecting its algal host from predation. Due to the presence of genes for vitamin biosynthesis, it might also provide the algal host with vitamins for growth and development. Some of these metabolic genes are biotechnologically important, as they could become a platform for bioengineering to generate various seaweed-derived substances sustainably, such as antibiofilm agents and vitamins, which are beneficial for human health.
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