Biotechnological Conversion of Grape Pomace to Poly(3-Hydroxybutyrate) by Moderately Thermophilic Bacterium Tepidimonas taiwanensis

Polyhydroxyalkanoates (PHA) are microbial polyesters that have recently come to the forefront of interest due to their biodegradability and production from renewable sources. A potential increase in competitiveness of PHA production process comes with a combination of the use of thermophilic bacteria with the mutual use of waste substrates. In this work, the thermophilic bacterium Tepidimonas taiwanensis LMG 22826 was identified as a promising PHA producer. The ability to produce PHA in T. taiwanensis was studied both on genotype and phenotype levels. The gene encoding the Class I PHA synthase, a crucial enzyme in PHA synthesis, was detected both by genome database search and by PCR. The microbial culture of T. taiwanensis was capable of efficient utilization of glucose and fructose. When cultivated on glucose as the only carbon source at 50°C, the PHA titers reached up to 3.55 g/L, and PHA content in cell dry mass was 65%. The preference of fructose and glucose opens the possibility to employ T. taiwanensis for PHA production on various food wastes rich in these abundant sugars. In this work, PHA production on grape pomace extracts was successfully tested.

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Extremophiles - Platform Strains for Sustainable Production of Polyhydroxyalkanoates

Materials Science Forum ◽

10.4028/www.scientific.net/msf.955.74 ◽

2019 ◽

Vol 955 ◽

pp. 74-79 ◽

Cited By ~ 1

Author(s):

Iva Pernicova ◽

Dan Kucera ◽

Ivana Novackova ◽

Juraj Vodicka ◽

Adriana Kovalcik ◽

...

Keyword(s):

Bacterial Biomass ◽

Pha Synthase ◽

Bacterial Cells ◽

Spent Coffee Grounds ◽

Production Potential ◽

Gene Encoding ◽

Biodegradable Polyesters ◽

Coffee Grounds ◽

Hypotonic Lysis ◽

Pha Production

Polyhydroxyalkanoates (PHA) are biodegradable polyesters, which are produced by various bacteria including numerous halophiles. Employment of halophilic strain for PHA production brings numerous benefits such as robustness of the process against contamination by ubiquitous mesophiles or possibility to isolate polymer from bacterial biomass via hypotonic lysis. In this work, we screened three moderate halophiles – Halomonas halophila, Halomonas organivorans and Halomonas salina for the presence of phaC gene encoding for PHA synthase and, subsequently, we have investigated their PHA production potential on various sugars. Among tested strains, H. organivorans demonstrated the extraordinary capacity of PHA production in particular on galactose and mannose since on these saccharides PHA content in dried bacterial cells reached 83 and 90 wt. % on mannose and galactose, respectively. Therefore, H. organivoras can be considered being promising PHA producing strain in particular suitable for the valorization of lignocellulose materials rich in galactomannans such as spent coffee grounds.

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Alterococcus agarolyticus, gen.nov., sp.nov., a halophilic thermophilic bacterium capable of agar degradation

Canadian Journal of Microbiology ◽

10.1139/w98-051 ◽

1998 ◽

Vol 44 (7) ◽

pp. 637-645 ◽

Cited By ~ 31

Author(s):

Wung Yang Shieh ◽

Wen Dar Jean

Keyword(s):

Organic Acids ◽

Hot Springs ◽

Thermophilic Bacteria ◽

Halophilic Bacteria ◽

Cellular Fatty Acid ◽

Thermophilic Bacterium ◽

Anaerobic Growth ◽

Moderately Thermophilic ◽

Degrading Bacteria ◽

Fermentative Bacteria

Five strains of facultatively anaerobic moderately thermophilic bacteria were isolated from two hot springs in the intertidal zone of Lutao, Taiwan. They produced extracellular agarase on agar medium, yielding reducing sugars and organic acids as the end products under either aerobic or anaerobic conditions. The growth temperature range was approximately 3858°C with an optimal temperature of about 48°C. The five strains tolerated a relatively narrow pH range from 7.0 to 8.5. They were Gram-negative halophiles growing optimally at 2.02.5% NaCl (ca. 0.340.43 M). They were capable of anaerobic growth by fermenting glucose and producing various organic acids such as butyrate, propionate, formate, lactate, and acetate. Cells grown in liquid medium were motile monotrichous cocci, normally 0.80.9 µm in diameter. They possessed saturated anteiso-15-carbon acid (anteiso-C15:0) as the most abundant cellular fatty acid (46.051.3 mo1%) and had G+C contents ranging from 65.5 to 67.0 mo1%. They are the first thermophiles found to degrade agar and also the first halophilic thermophilic bacteria known to be capable of both aerobic and anaerobic fermentative growth. These bacteria are considered to represent a new genus that we named Alterococcus, and Alterococcus agarolyticus is the type species. Key words: Alterococcus agarolyticus, thermophilic bacteria, halophilic bacteria, agar-degrading bacteria, fermentative bacteria.

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Effects of a Groundwater Heat Pump on Thermophilic Bacteria Activity

Water ◽

10.3390/w11102084 ◽

2019 ◽

Vol 11 (10) ◽

pp. 2084 ◽

Cited By ~ 1

Author(s):

Kim ◽

Lee

Keyword(s):

Heat Pump ◽

Power Plants ◽

Thermophilic Bacteria ◽

Thermophilic Bacterium ◽

Microbial Culture ◽

Temperature Dependent ◽

Psychrophilic Bacteria ◽

Groundwater Samples ◽

Groundwater Heat Pump ◽

Autochthonous Bacteria

Groundwater samples were collected from the tubular wells of a groundwater heat pump (GWHP), and the psychrophilic, mesophilic, and thermophilic bacteria inhabiting the collected groundwater were cultured and isolated. Using the isolated bacteria, we analyzed temperature-dependent changes in autochthonous bacteria based on the operation of the GWHP. Microbial culture identified eight species of bacteria: five species of thermophilic bacteria (Anoxybacillus tepidamans, Bacillus oceanisediminis, Deinococcus geothermalis, Effusibacillus pohliae, and Vulcaniibacterium thermophilum), one species of mesophilic bacteria (Lysobacter mobilis), and two species of psychrophilic bacteria (Paenibacillus elgii and Paenibacillus lautus). The results indicated A. tepidamans as the most dominant thermophilic bacterium in the study area. Notably, the Anoxybacillus genus was previous reported as a microorganism capable of creating deposits that clog above-ground wells and filters at geothermal power plants. Additionally, we found that on-site operation of the GWHP had a greater influence on the activity of thermophilic bacteria than on psychrophilic bacteria among autochthonous bacteria. These findings suggested that study of cultures of thermophilic bacteria might contribute to understanding the bio-clogging phenomena mediated by A. tepidamans in regard to GWHP-related thermal efficiency.

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Decomposition of Rice Chaff Using a Cocultivation System of Thermobifida fusca and Ureibacillus thermosphaericus

Proceedings ◽

10.3390/proceedings2020066031 ◽

2021 ◽

Vol 66 (1) ◽

pp. 31

Author(s):

Sachiko Nakamura ◽

Norio Kurosawa

Keyword(s):

Lignin Peroxidase ◽

Culture Medium ◽

Reducing Sugars ◽

Thermophilic Bacteria ◽

Thermobifida Fusca ◽

Bacterial Strains ◽

Basal Salt Medium ◽

Moderately Thermophilic ◽

Fuels And Chemicals ◽

Ureibacillus Thermosphaericus

Lignocellulosic biomass comprises cellulose, hemicellulose, and lignin and is a potential source of fuels and chemicals. Although this complex biomass is persistent, it can be cooperatively decomposed by a microbial consortium in nature. In this study, a coculture of the moderately thermophilic bacteria Thermobifida fusca and Ureibacillus thermosphaericus was used for biodegradation of rice chaff. The bacterial strains were incubated in modified Brock’s basal salt medium (pH 8.0) supplemented with yeast extract and rice chaff at 50 °C for 7 days. The concentration of reducing sugars and the enzymatic activities of laccase, lignin peroxidase, cellulase, and xylanase in the supernatant of the culture medium were measured every day. The concentrations of reducing sugars in solo cultures of T. fusca and U. thermosphaericus and a mixed culture of the two strains after 7 days of incubation were 0.047, 0.040, and 0.195 mg/mL, respectively, indicating that the decomposition of rice chaff was enhanced in the coculture. Based on the results, it is thought that the lignin surrounding the cellulose was decomposed by laccase and lignin peroxidase secreted from U. thermosphaericus, resulting in cellulose and hemicellulose in the rice chaff being easily decomposed by enzymes from T. fusca.

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Kinetics of medium-chain-length polyhydroxyalkanoate production by a novel isolate of Pseudomonas putida LS46

Canadian Journal of Microbiology ◽

10.1139/w2012-074 ◽

2012 ◽

Vol 58 (8) ◽

pp. 982-989 ◽

Cited By ~ 24

Author(s):

Parveen K. Sharma ◽

Jilagamazhi Fu ◽

Nazim Cicek ◽

Richard Sparling ◽

David B. Levin

Keyword(s):

Pseudomonas Putida ◽

Chain Length ◽

Saturated Fatty Acids ◽

Dry Mass ◽

Nucleotide Sequence Analysis ◽

Glucose Medium ◽

Medium Chain ◽

Medium Chain Length ◽

Pha Production ◽

Nitrogen Excess

Six bacteria that synthesize medium-chain-length polyhydroxyalkanoates (mcl-PHAs) were isolated from sewage sludge and hog barn wash and identified as strains of Pseudomonas and Comamonas by 16S rDNA gene sequencing. One isolate, Pseudomonas putida LS46, showed good PHA production (22% of cell dry mass) in glucose medium, and it was selected for further studies. While it is closely related to other P. putida strains (F1, KT2440, BIRD-1, GB-1, S16, and W619), P. putida LS46 was genetically distinct from these other strains on the basis of nucleotide sequence analysis of the cpn60 gene hypervariable region. PHA production was detected as early as 12 h in both nitrogen-limited and nitrogen-excess conditions. The increase in PHA production after 48 h was higher in nitrogen-limited cultures than in nitrogen-excess cultures. Pseudomonas putida LS46 produced mcl-PHAs when cultured with glucose, glycerol, or C6–C14 saturated fatty acids as carbon sources, and mcl-PHAs accounted for 56% of the cell dry mass when cells were batch cultured in medium containing 20 mmol/L octanoate. Although 3-hydroxydecanoate was the major mcl-PHA monomer (58.1–68.8 mol%) in P. putida LS46 cultured in glucose medium, 3-hydroxyoctanoate was the major monomer produced in octanoate medium (88 mol%).

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Identification of Poly(cis-1,4-Isoprene) Degradation Intermediates during Growth of Moderately Thermophilic Actinomycetes on Rubber and Cloning of a Functional lcp Homologue from Nocardia farcinica Strain E1

Applied and Environmental Microbiology ◽

10.1128/aem.72.5.3375-3382.2006 ◽

2006 ◽

Vol 72 (5) ◽

pp. 3375-3382 ◽

Cited By ~ 58

Author(s):

Ebaid M. A. Ibrahim ◽

Matthias Arenskï¿½tter ◽

Heinrich Luftmann ◽

Alexander Steinbï¿½chel

Keyword(s):

Thermophilic Bacteria ◽

Natural Rubber Latex ◽

Gel Permeation Chromatography ◽

Mycolic Acid ◽

Rrna Gene ◽

Ionization Mass ◽

Thermophilic Actinomycetes ◽

Moderately Thermophilic ◽

Nocardia Farcinica ◽

Different Strains

ABSTRACT The enrichment and isolation of thermophilic bacteria capable of rubber [poly(cis-1,4-isoprene)] degradation revealed eight different strains exhibiting both currently known strategies used by rubber-degrading mesophilic bacteria. Taxonomic characterization of these isolates by 16S rRNA gene sequence analysis demonstrated closest relationships to Actinomadura nitritigenes, Nocardia farcinica, and Thermomonospora curvata. While strains related to N. farcinica exhibited adhesive growth as described for mycolic acid-containing actinomycetes belonging to the genus Gordonia, strains related to A. nitritigenes and T. curvata formed translucent halos on natural rubber latex agar as described for several mycelium-forming actinomycetes. For all strains, optimum growth rates were observed at 50ï¿½C. The capability of rubber degradation was confirmed by mineralization experiments and by gel permeation chromatography (GPC). Intermediates resulting from early degradation steps were purified by preparative GPC, and their analysis by infrared spectroscopy revealed the occurrence of carbonyl carbon atoms. Staining with Schiff's reagent also revealed the presence of aldehyde groups in the intermediates. Bifunctional isoprenoid species terminated with a keto and aldehyde function were found by matrix-assisted laser desorption ionization-time-of-flight and electrospray ionization mass spectrometry analyses. Evidence was obtained that biodegradation of poly(cis-1,4-isoprene) is initiated by endocleavage, rather than by exocleavage. A gene (lcp) coding for a protein with high homology to Lcp (latex-clearing protein) from Streptomyces sp. strain K30 was identified in Nocardia farcinica E1. Streptomyces lividans TK23 expressing this Lcp homologue was able to cleave synthetic poly(cis-1,4-isoprene), confirming its involvement in initial polymer cleavage.

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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

Indonesian Journal of Biotechnology ◽

10.22146/ijbiotech.48272 ◽

2020 ◽

Vol 24 (2) ◽

pp. 105

Author(s):

Edvan Arifsaputra Suherman ◽

Maelita Ramdani Moeis ◽

Elvi Restiawaty

Keyword(s):

Amino Acids ◽

High Temperatures ◽

Hydrothermal Vent ◽

Animal Feed ◽

Thermophilic Bacteria ◽

Glycosyl Hydrolase ◽

Thermophilic Bacterium ◽

Lower Percentage ◽

Bacillus Species ◽

North Sulawesi

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.

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