Isolation Of Thermophilic Bacteria Research Articles

Isolation of Thermophilic Bacteria from Extreme Environments in Northern Chile.

The northern region of Chile boasts unique geographical features that support the emergence of geothermal effluents, salt lagoons, and coastal creeks. These extreme climate conditions create polyextreme habitats for microorganisms, particularly adapted to survive these harsh environments. These extremophilic microorganisms hold immense potential as a source of hydrolytic enzymes, among other biotechnological applications. In this study, we isolated 15 strains of aerobic thermophilic bacteria (45-70 °C) from sediment samples collected at five different ecological sites, including hot springs, geothermal fields, and lagoons in the Atacama Desert and Andes high planes. Analyses of the 16S rRNA gene sequences of the isolates showed a close genetic similarity (98-100%) with microorganisms of the genera Parageobacillus, Geobacillus, Anoxybacillus, and Aeribacillus. Notably, these thermophiles exhibited significant hydrolytic enzyme activity, particularly amylases, lipases, and proteases. These findings underscore the potential of using these thermophilic bacterial strains as an invaluable source of thermozymes with wide-ranging applications in diverse industries, such as detergent formulations, pharmaceutical processing, and food technology. This research highlights the ecological significance of these extreme environments in the Atacama Desert and Andes high plains, which serve as vital ecological niches housing extremophilic bacteria as a genetic source of relevant thermozymes, promising great potential for innovation in the biotechnology industry.

Identification and Study of Thermophilic Bacteria Found in the Sulfur-Infused Hot Springs of South Lampung

Thermophilic bacteria are microorganisms with the capability to produce a specific enzyme known as amylase, which exhibits resistance to heat and is referred to as thermostable. This study aims to isolate, characterize, and assess the enzyme-producing activity of bacteria from the hot springs of Way Belerang, Lampung, specifically amylase. The research procedures encompassed sample collection, isolation of thermophilic bacteria incubated in NA media for 48 hours, amylum testing with iodine after a 24-hour incubation in NA media containing starch/amylose, biochemical tests including TSIA, SIM, SC, urea test, sugar fermentation test, and catalase test. The results of the study yielded two isolates successfully isolated from the hot spring source, and Gram staining indicated that both isolates were Gram-negative bacteria. The assessment of bacterial activity in producing amylase was demonstrated by the presence of clear zones, with the largest bacterial diameter measuring 1.04 mm as measured using a vernier caliper. Characterization of the bacteria identified them as belonging to the Pseudomonas genus for isolates AB2 and AB3. Thus, the bacteria obtained in this study produce amylase enzymes that can be utilized in various industrial applications, including the food, beverage, textile, detergent, paper, pharmaceutical, and other sectors.

Effect of the Thermophilic Bacterial Biculture Consortium from Mudiak Sapan Hot Springs on Biofuel Production

Biofuels are fuels derived from biomass and consist of biogas, biodiesel and bioethanol. Bioethanol is a biofuel whose main ingredients are from plants and generally use microorganisms in the fermentation process. One way is to use thermophilic bacteria with the advantage of low contamination levels and better product quality. Bacteria that exist in nature are not only in a single form but also exist in the form of a consortium in which there are beneficial or detrimental bacterial interactions. Favorable bacterial interactions indicate that the bacteria are compatible. Using a consortium with compatible bacteria gives better results than using a single bacteria. This study aims to determine the compatibility and effect of a consortium of thermophilic bacteria from Mudiak Sapan hot springs on biofuel yields. This research is a descriptive research. To test the cooperation between isolates, a compatibility test was carried out using the disk diffusion method. Then the biculture consortium isolates of thermophilic bacteria were fermented in liquid TMM medium (Thermophilic Minimum Media) and the bioethanol content was measured using a distillation apparatus. The results of this study showed that the six pairs of compatible consortium and MS 9-12 consortium produced the highest bioethanol, namely 1.0003%. Then the biculture consortium isolates of thermophilic bacteria were fermented in liquid TMM medium (Thermophilic Minimum Media) and the bioethanol content was measured using a distillation apparatus. The results of this study showed that the six pairs of compatible consortium and MS 9-12 consortium produced the highest bioethanol, namely 1.0003%. Then the biculture consortium isolates of thermophilic bacteria were fermented in liquid TMM medium (Thermophilic Minimum Media) and the bioethanol content was measured using a distillation apparatus. The results of this study showed that the six pairs of compatible consortium and MS 9-12 consortium produced the highest bioethanol, namely 1.0003%.

Isolation, Screening and Partial Characterization of Thermophilic Bacteria Producing Protease from Bukik Gadang Hot Springs, Solok Regency

Research about the isolation, screening and partial characterization of thermophilic bacteria that produce protease from Bukik Gadang hot springs, Solok Regency, was carried out at the Basic Biology Laboratory, UPT Basic and Central Laboratory, Andalas University in January - April 2022. This study aims to obtain isolates of thermophilic bacteria that indicated protease production and to determine the partial characterization of thermophilic bacteria that indicated protease production from Bukik Gadang hot springs, Solok Regency. This study used a survey method and purposive sampling technique. The results of this study obtained 18 isolates from 20 isolates of thermophilic bacteria which indicated protease production, with one isolate of TPBG-03 having the potential to produce protease. Partial characterization by 10 isolates of thermophilic bacteria indicated protease production was generally Gram positive, bacillus cell shape, spore-forming, catalase positive and motile.Keywords - Hot Springs, Isolation, Protease, Screening, Thermophilic Bacteria

Isolation of Thermophilic Bacteria and Investigation of Their Microplastic Degradation Ability Using Polyethylene Polymers.

Microplastics (MPs) pose potential public health challenges because of their widespread occurrences in all environmental compartments. While most studies have focused on the occurrence fate of microplastics in wastewater treatment systems, the biodegradation of microplastics in wastewater is generally little understood. Therefore, we used two Gram-positive and thermophilic bacteria, called strain ST3 and ST6, which were identified by morphological, biochemical, physiological, and molecular analyses, to assess the growth and biodegradation potential of two different sized (50 and 150 m) polyethylene particles. The degradation was monitored based on structural and surface morphological changes. According to 16S rRNA analyses, ST3 and ST6 were identified as Anoxybacillus flavithermus ST3 and Anoxybacillus sp. ST6, respectively. The occurrence of cracks, holes, and dimensional changes was detected by scanning electron microscopy. Moreover, critical characteristic absorption band formation and modifications were determined by Fourier transform infrared spectroscopy. In addition to these, it was found that Anoxybacillus flavithermus ST3 and Anoxybacillus sp. ST6 produced high level of alpha-Amylase. These results showed that thermophilic bacteria are capable of the biodegradation of microplastics and production of alpha-Amylase.

ISOLASI DAN IDENTIFIKASI BAKTERI TERMOFILIK PENGHASIL AMILASE DARI SUMBER AIR PANAS SIDEBUK-DEBUK SUMATERA UTARA

Hot springs or hot springs are springs that are produced as a result of the release of groundwater from the earth's crust after experiencing geothermal heating. Indonesia is one of the most active tectonic regions in the world with more than 70 active volcanoes, and has many geothermal areas. Hot springs are a suitable growth medium for thermophilic bacteria. Thermophilic bacteria are a group of bacteria that adapt to high-temperature environmental conditions, with temperatures ranging from 50°C-75°C. The natural habitat of thermophilic bacteria is widespread throughout the earth's surface, including in hot springs, volcanic craters or volcanic areas. One of the hot springs in North Sumatra province is sidebuk-debuk, which is located in Bandar Baru sub-district. is a hot spring, which has a temperature between 50°C – 75°C. At this time thermophilic bacteria are studied and researched intensively for reasons of developing basic research and biotechnology applications. Thermophilic bacteria have the potential to produce heat-resistant/ thermostable enzymes and can be used in industry, waste treatment processes and mineral weathering. Amylase is an important enzyme that is widely used in industrial applications through synthesis reactions and hydrolysis reactions, accounting for almost 65% of the total sales of enzymes in the world. Researchers are interested in knowing more about the role of thermostable enzymes in modern biotechnology today, therefore it is necessary to conduct research aimed at isolating and identifying local isolates of thermophilic bacteria as potential sources of enzymes, especially in North Sumatra. From this study, 19 isolates of Bacillus sp bacteria were obtained from Sidebuk-debuk hot springs based on phenotypic characters by looking at macroscopic, microscopic and biochemical characteristics, with 9 isolates of thermophilic bacteria that have the potential to produce amylase as indicated by the presence of a clear zone around the bacterial colonies.Keywords: Hot Springs, Bacillus sp Bacteria,

Thermophilic bacteria isolated from Mount Merapi, Java, Indonesia as a potential lead bioremediation agent

Abstract. Rakhmawati A, Wahyuni ET, Yuwono T. 2021. Thermophilic bacteria isolated from Mount Merapi, Java, Indonesia as a potential lead bioremediation agent. Biodiversitas 22: 3101-3110. Contamination by lead (Pb) has become a serious health and environmental problem, that has to be urgently prevented. Bioremediation is one of the environmentally friendly methods for eliminating Pb from the contaminated environment. This study was aimed to investigate the potency of lead-tolerant thermophilic bacteria isolated from solfatara Mount Merapi, Indonesia. A total of 340 isolates of thermophilic bacteria were screened for lead tolerance at 55 °C. Five bacterial isolates were found to show tolerance to 100 mg/L Pb (II), and then were further evaluated and identified as Aeribacillus pallidus strains MRP112, MRP148, MRP272, MRP278, and MRP280 based on 16S rRNA gene sequences analysis. Among the five isolates, A. pallidus MRP 280 showed highest activity in removing Pb at pH 6, 55°C for 24 h. The analysis of scanning and transmission electron microscopy, biofilm formation, and siderophore production, demonstrated that lead tolerance of A. pallidus MRP 280 strain was also accompanied by morphological changes, bioaccumulation, biosorption, biofilm, and siderophore assembly. In conclusion A. pallidus MRP 280 was demonstrated as one of the most potential bacterial strains, can be recommended as an agent for high-temperature lead bioremediation.

Potential enzyme activity of thermophilic bacteria from hot spring in Egypt

This study aims to isolate and identify thermophilic hydrolytic bacteria from hot spring in South Sinai, Egypt for several industrial applications. In this work, 29 bacterial isolates from hot spring in Egypt were isolated and screened for the production of three thermozymes (amylase, cellulase and protease) at different high temperatures. Fifteen isolates were amylase producer, twenty-one were cellulase producer and twelve isolates were protease producer at different high temperatures. Ten bacterial isolates (34.5%) produced the three extracellular enzymes. All bacterial isolates were identified phenotypically as Bacillus spp. This study concludes that hot springs in Egypt is a source for the isolation of thermophilic bacteria producing thermostable enzymes.

Isolation and Characterization of Thermophilic Bacteria as Cellulolytic Enzyme Producer from the Hot Spring of Ie Seuum Aceh Besar, Indonesia

Cellulase enzymes can be isolated from thermophile bacteria obtained from the hot spring Ie Seuum, Aceh Besar. This research aimed to recover and characterize the isolates morphologically and biochemically followed by determination of the thermophile bacterial isolates potential as cellulolytic enzyme producers,. The sampling method in this research was conducted by a purposive sampling at temperature of 70 oC, 60 oC and 50 oC. Isolation of thermophilic bacteria was carried out on nutrient agar (NA) media. There were four isolates of thermophilic bacteria isolated recovered at 70 oC, five isolates at 60 oC, and seven isolates at 50 oC. Of the 18 isolates obtained, 15 of them were able to produce cellulase enzymes. Cellulase enzyme production can be determined by the presence of clear zones around bacterial colonies on CMC media after addition of 1% congo red drops and wash with 1 M NaCl. The highest five Cellulolytic Index (CI) values ​​were obtained from isolates ISB75; ISB64; ISB52; ISB54; ISB56 that were 1.23; 2.22; 1.39; 1.59; 1.10, respectively. Biochemical tests carried out on 5 isolates with the highest cellulolytic index values showed that the bacterial isolate were suspected to be from the genera of Bacillus sp.

Cloning and in silico study of an endoglucanase from a thermophilic bacterium isolated from a hydrothermal vent of West Kawio, Sangihe‐Talaud waters, North Sulawesi, Indonesia

Endoglucanase is used in industries that apply high temperatures, such as bioethanol, detergent, paper, and animal feed industries. Most available endoglucanases have very low stability at high temperatures. Therefore, this study aimed to identfy a new thermostable endoglucanase that is able to maintain its actvity at high temperatures. Five isolates of thermophilic bacteria were previously isolated from the hydrothermal vent of West Kawio, Indonesia. Among them, the DSI2 isolate showed the highest endoglucanase actvity, and was identfed and named as Bacillus safensis DSI2. The EgDSI2 gene was cloned from B. safensis DSI2. EgDSI2 is 1851 bp long encoding a protein of 616 amino acids. The encoded protein, EgDSI2, has high sequence identty to other B. safensis endoglucanases and was predicted with the Compute pI/Mw tool to be 69.41 kDa. EgDSI2 was high in hydrophobic amino acids. The enzyme had higher percentage of Ala andPro, and lower percentage of Gly compared to thermolabile endoglucanases from two Bacillus species. EgDSI2 harbored a catalytc domain belonging to glycosyl hydrolase family 9 (GH9) and a type 3 cellulose‐binding domain (CBM3). Propertes of endoglucanases with GH9‐CBM3 modular organizaton include actvity over a wide pH range, high optmum temperature, and thermostablity. Therefore, EgDSI2 has potental applicatons in the industries.

Inulinase Activity of Thermophilic Bacteria isolated from Hot Springs of Penen Village, North Sumatera, Indonesia

Inulinase is an enzyme that catalyzes inulin to produce D-fructose as the end product which is considered as safe alternative sweetener to sucrose. This enzyme can be extracted from plants and vegetables but it is difficult to obtain it in high quantity and the production cost also high. The main problem in utilizing this enzyme in industrial is the instability against fluctuation of temperature that usually occurs in industrial process. Therefore thermophilic microbial inulinase has great potential for these industrial needs. The aim of this study was to isolate and characterize the thermophilic bacteria that possess the inulinase activity from hot springs in Penen Village, North Sumatera. To screening the inulinase producing bacteria, the modified Czapex Dox agar supplemented with 1,5% inulin powder was used. The temperature used for incubation is 48 °C. The clear zone surrounds the bacteria after Lugol’s iodine treatment on medium indicated the nucleolytic activity. As 4 isolates of thermophilic bacteria namely UTMP 11, UTMP 14, UTMP 15 and UTMP 17 showed the inulolytic activity. The biochemical characterization of these isolates showed negative in catalase test, negative on citrate test, non-motil, negative Gram staining, and rod-shaped morphology. Molecularly, this bacteria identified as Bacillus subtilis strain JCM 1465 and Bacillus tequilensis strain 10b. From the result, these isolates have potential as source of thermophilic inulinase production in food industry for the production of fructose.

Molecular Barcoding Based 16S rRNA Gene of Thermophilic Bacteria from Vulcanic Sites, Linow Lake, Tomohon

Thermophilic bacteria live at temperatures above 450 C. Many investigations focused on their potential as sources of highly active enzymes ‘termostable enzyme’ and other products such as antibiotics and compatible solutes. Lake Linow is an active volcanic lake located in Tomohon City, North Sulawesi, Indonesia. Lake Linow becomes the habitat of thermophilic bacteria. A study has been conducted to obtain isolates of thermophilic bacteria and to identifikasi berdasarkan gen 16 s RNA. Bacterial DNA extraction procedure using the Presto TM Mini gDNA Bacteria Kit Geneaid protocol, with modifications. Amplification of 16s RNA gene using PCR method. Visualization of 16 s RNA amplicon genes with automatic electrophoresis capiler Qiaxel, Qiagen. Sequencing was carried out using Singapore's First BASE Sequencing service. The results showed that IL2 isolates and IL3 isolates could live up to 700C. Alignment analysis results using NCAST BLBI IL2 isolates showed 99% similarity with Bacillus thuringiensis strain H2682 (accession number CP009720.1). While isolate of IL3 thermophilic bacteria showed 94% similarity with Bacillus licheniformis strain 14DA11 (accession number CP023168.1). The results of phylogeny reconstruction with neighbor joining method, gene sequence 16S rRNA isolate IL2 showed the closest relation with Bacillus thuringiensis strain HD1011 (accession number CP009335.1). While IL3 isolate showed the closest relation with Bacillus licheniformis strain 14DA11 (accession number CP023168.1).

Isolasi dan Identifikasi Bakteri Termofilik dari Sumber Air Panas di Moso Distrik Muara Tami Kota Jayapura Provinsi Papua

Thermophilic bacteria are bacteria that are able to survive in environmental conditions with high temperatures. Thermophilic bacteria are one of the important sources of thermostable enzymes that can be isolated from geothermal environments such as hot springs with temperatures ranging from 50–80 oC. Aims of the study to isolate and identify local isolates of thermophilic bacteria from hot springs in Moso Muara Tami District, Papua. The results showed that 7 isolates (A1, A2, A3, A4, B1, B2, and B3) that were isolated and identified based on phenotypic characters were included in the genus Bacillus.  Key words: Thermophilic, characterization, isolation, identification, phenotypic.

Isolation of Thermophilic Bacteria from Bora Hot Springs in Central Sulawesi

Thermophilic bacteria can survive at high temperature, in which hot spring is one of its habitats. Indonesia has many hot springs with potential as a habitat for thermophilic bacteria. The purpose of this study was to obtain isolates thermophilic bacteria from Bora hotspring located in Central Sulawesi. This study applied a descriptive-observational study design, characteristics of bacterial properties identified using conventional methods according to the Bergey’s Manual of Determinative Bacteriology. The study was conducted in 3 stages. The first stage was bacteria cultivation on the appropriate media, followed by stage of isolated and the last stage by identified characteristics of thermophilic bacteria which included microscopic and macroscopic morphology, Physiological and biochemistry test. The results of the isolates indicted 4 representative isolated of thermophilic bacteria from Bora Hot Spring namely TM022, TM023, TM024, TM026. The bacteria isolates obtained were bacillus, coccus and Gram negative and positive, while the physiological test of all isolates were able to grow and showed changes in the medium. This study is useful in providing characteristic of indigenous thermophilic bacteria isolates that produces thermostable enzymes.

Potency of Amylase-producing Bacteria and Optimization Amylase Activities

Enzymes are capable to act as biocatalyst for a wide variety of chemical reactions. Amylase have potential biotechnological applications in a wide range of industrial processes and account for nearly 30% of the world’s enzyme market. Amylase are extracellular enzymes that catalyze the hydrolysis of internal α-1,4-glycosidic linkages in starch to dextrin, and other small carbohydrate molecules constituted of glucose units. Although enzymes are produced from animal and plant sources, the microbial sources are generally the most suitable for commercial applications. Bacteria from hot springs is widely used as a source of various enzymes, such as amylase. But the amount of amylase-producing bacteria is still very limited. Therefore it is necessary to search sources of amylase-producing bacteria new, such as from hot springs Pariangan. The purpose of this study was to isolation of amylase-producing bacteria from Pariangan hot spring, West Sumatera and amylase activity optimization. The results were obtained 12 isolates of thermophilic bacteria and 5 isolates of amyalse-producing bacteria with the largest amylolytic index of 3.38 mm. The highest amylase activity was obtained at 50°C and pH 7.5.

Screening and characterization of thermo-active enzymes of biotechnological interest produced by thermophilic Bacillus isolated from hot springs in Tunisia.

As part of the contribution to the global efforts in research of thermostable enzymes being of industrial interest, we focus on the isolation of thermophilic bacteria from Tunisian hot springs. Among the collection of 161 strains of thermophilic Bacillus isolated from different samples of thermal water in Tunisia, 20% are capable of growing at 100°C and the rest grow at 70°C or above. Preliminary activity tests on media supplemented with enzyme-substrates confirmed that 35 strains produced amylases, 37 - proteases, 43 - cellulases, 31 - xylanases and 37 - mannanases. The study of the effect of temperature on enzyme activity led to determination of the optimal temperatures of activities that vary between 60 and 100°C. Several enzymes were active at high temperatures (80, 90 and 100°C) and kept their activity even at 110°C. Several isolated strains producing enzymes with high optimal temperatures of activity were described for the first time in this study. Both strains B62 and B120 are producers of amylase, protease, cellulase, xylanase, and mannanase. The sequencing of 16S DNA identified isolated strains as Geobacillus kaustophillus, Aeribacillus pallidus, Geobacillus galactosidasus and Geobacillus toebii.

Isolation, characterization and molecular identification of protease producing bacteria from Tashkooh mountain located in Ahvaz, Iran

Thermophilic microorganisms have gained worldwide importance due to their remarkable potential to produce thermostable and thermoactive enzymes that have wide applications in pharmaceuticals and industries. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are very important in terms of discovering new industrial enzymes. The aim of this research was therefore the isolation of protease producing thermophilic bacteria from Tashkooh or firing mount located in Ahvaz, Iran. 8 bacterial isolates were screened. These strains were examined for the existence of extracellular protease activity. All 8 isolated bacteria showed proteolytic activity on nutrient agar with skim-milk. 3 bacteria showed their optimum growth at alkaline pH and grew maximally at different temperature in the thermophilic range and had proteolytic activity at pH 11 in 70?C and pH 9 in 55?C.The best carbon source for proteolytic activity was starch. After performing some phenotypic tests determined that all the isolates were Gram positive, endospore forming rods, aerobic, capable to produce catalase, amylase and gelatinase enzymes and they were identified as Bacillus sp. The best isolated bacteria was identified molecularly with the aid of 16S rRNA sequencing and data revealed that the Bacillus subtilis strain G-13 (GenBank accession No. KJ139434.1).The investigation confirmed that the isolate to be a true thermophile and could be a source of thermostable protease which can be exploited for pharmaceutical and industrials applications.DOI: http://dx.doi.org/10.3126/ijls.v9i2.12054 International Journal of Life Sciences 9 (2) : 2015; 39-42

Estimation of extracellular lipase enzyme produced by thermophilic bacillus sp. isolated from arid and semi-arid region of Rajasthan, India

AbstractThermophilic organisms can be defined as microorganisms which are adapted to live at high temperatures. The enzymes produce by thermophilic bacteria are capable of catalyzing biochemical reactions at high temperatures. Thermophilic bacteria are able to produce thermostable lipase enzymes capable of degradation of lipid at temperatures higher than those of mesophilic bacteria. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are quite useful in terms of discovering thermophilic lipase enzymes. Due to great temperature fluctuation in hot arid and semi-arid regions of Rajasthan, this area could serve as a good source for new thermophilic lipase producing bacteria with novel industrially important properties. The main objective of this research is the isolation and estimation of industrially important thermophilic lipase enzyme produced by thermophilic bacteria, isolated from arid and semi-arid region of Rajasthan. For this research purpose, soil samples were collected from Churu, Sikar and Jhunjunu regions of Rajasthan. A total of 16 bacterial strains were isolated, and among these bacterial isolates only two thermophilic lipase producing bacteria were identified. The thermophilic lipase enzyme was estimated by qualitative and quantitative experiments. The isolate was identified as Bacillus sp. by microscopic, biochemical and molecular characterization. The optimum enzyme activity was observed at pH 8, temperature 60°C and 5% salt concentration at 24 hrs time duration. Lipases are useful in a variety of biotechnological fields such as food and dairy (cheese ripening, flavour development), detergent, pharmaceutical (naproxen, ibuprofen), agrochemical (insecticide, pesticide) and oleochemical (fat and oil hydrolysis, biosurfactant synthesis) industries. Lipases can be further used in many newer areas where they can serve as potential biocatalysts.

Population dynamics and extracellular enzymes activity of mesophilic and thermophilic bacteria isolated from semi-arid soil of Northeastern Brazil

The dynamics of mesophilic and thermophilic bacterial population was studied in the bulk soil and rhizosphere of the grass Aristida adscensionis L. in a caatinga of the semi-arid Brazilian Northeast. Mesophilic heterotrophic bacteria, sporeforming bacteria and actinomycetes were significantly more abundant than thermophiles, and their counts were 106 - 107 CFU g-1 dry soil, showing weak fluctuations over the one-year study period. Thermophiles were below 106 CFU g-1 dry soil and, in general, demonstrated stronger fluctuations over the studied period. Among them, thermophilic actinomycetes showed the greatest decline during the dry season. No significant differences in microbial number were observed between bulk and rhizosphere soil. Mesophilic bacteria were represented mainly by Gram positive, filamentous organisms (actinomycetes) and by Gram positive, endospore-forming, rod-shaped organisms. All isolates of thermophilic bacteria were Gram positive, endospore-forming rods. Most of mesophilic isolates produced amylases and proteases, and 38% of isolates produced all studied extracellular enzymes: amylases, proteases, cellulases and chitinases. Among thermophilic bacteria, all isolates showed proteolytic activity and 77% produced amylases, but no one produced cellulases or chitinases.

Two new thermostable type II restriction endonucleases from Thermus aquaticus: TatI and TauI, which recognize the novel nucleotide sequences 5'-W (downward arrow)GTACW-3' and 5'-GCSG (downward arrow)C-3' respectively.

One hundred and forty isolates of thermophilic bacteria from the genus Thermus were screened for the presence of restriction endonuclease activity. Thermostable isoschizomers of restriction endonucleases, such as AceIII, BbvI, BglI, BsePI, FnuDII, HgiAI, MaeII, MboI, MseI, PvuII, StuI, TaqI, Tsp4CI, TspEI, XhoI and XmaIII, were isolated. Two restriction enzymes, TatI and TauI, recognizing novel degenerate sequences 5'-W (downward arrow)GTACW-3' and 5'-GCSG (downward arrow)C-3' respectively were partially purified and the recognition and cleavage sites were determined.