• Journal of Microbiology and Biotechnology
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• v.34 no.9
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• pp.1867-1875
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• 2024

Identification of the biochemical metabolic pathway for lignin decomposition and the responsible degradative enzymes is needed for the effective biotechnological valorization of lignin to renewable chemical products. In this study, we investigated the decomposition of kraft lignin by the soil bacterium Pseudomonas kribbensis CHA-19, a strain that can utilize kraft lignin and its main degradation metabolite, vanillic acid, as growth substrates. Gel permeation chromatography revealed that CHA-19 decomposed polymeric lignin and degraded dehydrodivanillin (a representative lignin model compound); however, the degradative enzyme(s) and mechanism were not identified. Quantitative polymerase chain reaction with mRNAs from CHA-19 cells induced in the presence of lignin showed that the putative genes coding for two laccase-like multicopper oxidases (LMCOs) and three dye-decolorizing peroxidases (DyPs) were upregulated by 2.0- to 7.9-fold compared with glucose-induced cells, which indicates possible cooperation with multiple enzymes for lignin decomposition. Computational homology analysis of the protein sequences of LMCOs and DyPs also predicted their roles in lignin decomposition. Based on the above data, CHA-19 appears to initiate oxidative lignin decomposition using multifunctional LMCOs and DyPs, producing smaller metabolites such as vanillic acid, which is further degraded via ortho- and meta-ring cleavage pathways. This study not only helps to better understand the role of bacteria in lignin decomposition and thus in terrestrial ecosystems, but also expands the biocatalytic toolbox with new bacterial cells and their degradative enzymes for lignin valorization.

• Journal of Food Hygiene and Safety
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• v.5 no.3
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• pp.131-138
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• 1990

Aflatoxins is a chemically diverse group of toxic secondary metabolites that are produced by fungi and often occur in agricultural commodities. Because of their wide range of toxic effects, Aflatoxins cause severe economic losses to farmers and livestock producers and pose a health to human consuming contaminated foods. Long term prospects for biotechnological control of Aflatoxins require elucidation of the specific steps and regulation of their biosynthetic pathways . Aflatoxin determinations can be approached many ways. It is essential to safely handle all experimental materials associated with aflatoxin analysis or aflatoxigenic fungi Visual screening of suspect samples, base on the presence of conidial head of the aspergillus flavus group, and screening samples for the presence of bright greenish yellow flourescence are not chemical tests and such screening techniques may allow aflactoxin contaminated lots into commerce. Microcolumn screening procedures should always be used in conjunction with a quantitative method. Several thin layer chromatography(TLC) and high performance liquid chromatography(HPLC) methods are suitable for quantitation and are in general use. Immunochemical Methods such as the ELISA or affinity column chromatography methods are being rapidly developed. The chemical and immunochemical methods can be reliable if care is taken, using suitable controls and personnel that are well trained . All analytical laboratories should stress safety and include suitable analytical validation procedure. Especially a worldwide enquiry was undertaken in recent to obtain up-to-date information about aflatoxin legislation in as many countries of the world as possible. The information concerns aflatoxin in foodstuffs. aflatoxin MI in dairy products, aflatoxins in animal feedstuffs. Limits and regulations for aflatoxin have been expended in recent with more countries having legislation on subject, more products, and more aflatoxins covered by this legislation.

• Hwankyungkyoyuk
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• v.23 no.1
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• pp.99-111
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• 2010

The purpose of this study was to investigate high school students' perceptions of biotechnology. Participants in this study were 9th and 10th grade students who were enrolled in high schools in Gyeonggi Province. The survey instrument used in this study was a 26-item questionnaire that was designed to measure students' perceptions regarding biotechnology. The study revealed that students' perceptions were positive toward the use of biotechnology on biological objects such as plant, grain and microbes. However, their perceptions were negative toward the use of biotechnology on humans and animals. Male students' perceptions were more positive than female students and there were significant differences between male and female students(p<.01). The study also revealed that male and female students had positive perceptions about the use of biotechnology in the development of beneficial products. However, male students' perceptions were more positive than female students(p<.01). Female students' perceptions were slightly more negative than males and they indicated a measure of caution in the development of beneficial products using biotechnology. Regarding the reliability of biotechnological information acquired from food companies, TV broadcasters, and entertainers, male and female students tended to be highly negative. Students perceived that environmental, religious and ethical issues did not affect the use of biotechnology when asked the effect of these factors on the use of biotechnology. They perceived that food safety and genetic factors of microbes did affect the use of biotechnology. Thus, the study findings suggest that teaching and learning strategies based on the differences of perceptions between male and female students of this study be established and the use of media, development of teaching method and materials be promoted in order to enhance student's performance in environmental education.

• Journal of Marine Bioscience and Biotechnology
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• v.12 no.1
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• pp.11-19
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• 2020

This study aims to review the progress of biotechnological applications of sea cucumber and presents fundamental validation data on a range of methods and applications to determine the most promising fields for future exploitation and application. Sea cucumbers from different regions, subjected to numerous industrially applicable drying methods, were evaluated for their biochemical composition. The study highlights the nutritional excellence of Jeju red sea cucumber, particularly the high amino acid and fatty acid composition of its intestinal organs and the possibility of the industrial utilization of these organs. In addition, an efficient extraction method to extract collagen and chondroitin sulfate from red sea cucumber was established. The extracted collagen and chondroitin sulfate can be used as commercially valid biomaterials. Accordingly, the red sea cucumber was confirmed as a valuable source of raw food material of varied functionality and bioactivity. Upon conducting an analysis of patents thus far relating to the sea cucumber, it was found that the associated technology was limited to some parts of primary and secondary industries. Therefore, there is a clear need for the strategic development of technologies to produce specialized functional products from sea cucumber's raw materials, with a view to promote other related industries.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.712-717
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• 2010

Cytochrome P450 enzymes (P450s) are involved in the synthesis of a wide variety of valuable products and in the degradation of numerous toxic compounds. The P450 BM3 (CYP102A1) from Bacillus megaterium was the first P450 discovered to be fused to its redox partner, a mammalian-like diflavin reductase. Here, we report the development of a whole-cell biocatalyst using ice-nucleation protein (Inp) from Pseudomonas syringae to display a hemeand diflavin-containing oxidoreductase, P450 BM3 (a single, 119-kDa polypeptide with domains of both an oxygenase and a reductase) on the surface of Escherichia coli. The surface localization and functionality of the fusion protein containing P450 BM3 were verified by flow cytometry and measurement of enzymatic activities. The results of this study comprise the first report of microbial cell-surface display of a heme- and diflavin-containing enzyme. This system should allow us to select and develop oxidoreductases containing heme and/or flavins into practically useful whole-cell biocatalysts for extensive biotechnological applications, including selective synthesis of new chemicals and pharmaceuticals, bioconversion, bioremediation, live vaccine development, and biochip development.

• Journal of Microbiology
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• v.44 no.4
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• pp.396-402
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• 2006

[ ${\beta}-Galactosidase$ ] is extensively employed in the manufacture of dairy products, including lactose-reduced milk. Here, we have isolated two gram-negative and rod-shaped coldadapted bacteria, BS 1 and HS 39. These strains were able to break down lactose at low temperatures. Although two isolates were found to grow well at $10^{\circ}C$, the BS 1 strain was unable to grow at $37^{\circ}C$. Another strain, HS-39, evidenced retarded growth at $37^{\circ}C$. The biochemical characteristics and the results of 16S rDNA sequencing identified the BS 1 isolate as Rahnella aquatilis, and showed that the HS 39 strain belonged to genus Buttiauxella. Whereas the R. aquatilis BS 1 strain generated maximal quantities of ${\beta}-galactosidase$ when incubated for 60h at $10^{\circ}C$, Buttiauxella sp. HS-39 generated ${\beta}-galactosidase$ earlier, and at slightly lower levels, than R. aquatilis BS 1. The optimum temperature for ${\beta}-galactosidase$ was $30^{\circ}C$ for R. aquatilis BS-1, and was $45^{\circ}C$ for Buttiauxella sp. HS-39, thereby indicating that R. aquatilis BS-1 was able to generate a cold-adaptive enzyme. These two cold-adapted strains, and most notably the ${\beta}-galactosidase$ from each isolate, might prove useful in some biotechnological applications.

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.459-466
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• 2013

The lactic acid bacterium Streptococcus thermophilus is widely used as a starter culture for the production of dairy products. Whole-genome sequencing is expected to utilize the genetic basis behind the metabolic functioning of lactic acid bacterium (LAB), for development of their use in biotechnological and probiotic applications. We sequenced the whole genome of Streptococcus thermophilus MTCC 5461, the strain isolated from a curd source, by 454 GS-FLX titanium and Ion Torrent PGM. We performed comparative genome analysis using the local BLAST and RDP for 16S rDNA comparison and by the RAST server for functional comparison against the published genome sequence of Streptococcus thermophilus CNRZ 1066. The whole genome size of S. thermophilus MTCC 5461 is of 1.73Mb size with a GC content of 39.3%. Streptococcal virulence-related genes are either inactivated or absent in the strain. The genome possesses coding sequences for features important for a probiotic organism such as adhesion, acid tolerance, bacteriocin production, and lactose utilization, which was found to be conserved among the strains MTCC 5461 and CNRZ 1066. Biochemical analysis revealed the utilization of 17 sugars by the bacterium, where the presence of genes encoding enzymes involved in metabolism for 16 of these 17 sugars were confirmed in the genome. This study supports the facts that the strain MTCC 5461 is nonpathogenic and harbors essential features that can be exploited for its probiotic potential.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1670-1679
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• 2020

The substantial use of fungal enzymes to degrade lignocellulosic plant biomass has widely been attributed to the extensive requirement of powerful enzyme-producing fungal strains. In this study, a two-step screening procedure for finding cellulolytic fungi, involving a miniaturized culture method with shake-flask fermentation, was proposed and demonstrated. We isolated 297 fungal strains from several cellulose-containing samples found in two different locations in Thailand. By using this screening strategy, we then selected 9 fungal strains based on their potential for cellulase production. Through sequence-based identification of these fungal isolates, 4 species in 4 genera were identified: Aspergillus terreus (3 strains: AG466, AG438 and AG499), Penicillium oxalicum (4 strains: AG452, AG496, AG498 and AG559), Talaromyces siamensis (1 strain: AG548) and Trichoderma afroharzianum (1 strain: AG500). After examining their lignocellulose degradation capacity, our data showed that P. oxalicum AG452 exhibited the highest glucose yield after saccharification of pretreated sugarcane trash, cassava pulp and coffee silverskin. In addition, Ta. siamensis AG548 produced the highest glucose yield after hydrolysis of pretreated sugarcane bagasse. Our study demonstrated that the proposed two-step screening strategy can be further applied for discovering potential cellulolytic fungi isolated from various environmental samples. Meanwhile, the fungal strains isolated in this study will prove useful in the bioconversion of agricultural lignocellulosic residues into valuable biotechnological products.

• Korean Journal of Microbiology
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• v.55 no.1
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• pp.9-16
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• 2019

Acetoin is generated by numerous microorganisms using ${\alpha}$-acetolactate decarboxylase (ALDC) to prevent overacidification of cells and their environment and to store remaining energy. Because acetoin has been used as a safe flavor enhancer in food products, industries have been interested in biotechnological production of acetoin using ALDC. ALDC is a metal-dependent enzyme that produces acetoin from ${\alpha}$-acetolactate through decarboxylation reaction. Here, we report the crystal structure of ALDC from Bacillus subtilis subspecies spizizenii (bssALDC) at $1.7{\AA}$ resolution. bssALDC folds into a two-domain ${\alpha}/{\beta}$ structure where two ${\beta}$-sheets form a central core. bssALDC assembles into a dimer through central hydrophobic interactions and peripheral hydrophilic interactions. bssALDC coordinates a zinc ion using three histidine residues and three water molecules. Based on comparative analyses of ALDC structures and sequences, we propose that the active site of bssALDC includes the zinc ion and its neighboring bssALDC residues.

• Journal of the Korean Society of Industry Convergence
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• v.22 no.1
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• pp.61-69
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• 2019

All the parts of lotus, including the seed, rhizome, leaf, stalk, petal, anther, pericarp, and fruit receptacle, have been used in traditional medicine system as a health beneficial supplement. However the most usually used from lotus plant is only the root. Therefore in this study, it will be discussed more the utilization of other parts of the lotus, namely the flower of lotus. The petals and stamens of lotus actually are also rich in bioactive components such as flavonoids and alkaloids, are used in the treatment of tissue inflammation, cancer, skin disease, and also for us as antidotes. One of the biotechnological process that can be used to improve the nutritional content, sensory, and also antioxidant activities is fermentation process. The final product desired from the fermentation process in this study is vinegar. The microbial strain powder used is Uinkin fermented powder with three variations of fermentation. The variations given in this study were initial sugar 32%, 24%, and 14% with the same fermentation temperature, $35^{\circ}C$ for 3 months. The results obtained showed that the pH value and sugar content of products during the fermentation process were decreasing during the fermentation process, with total polyphenol content of $283.7{\pm}97.6mg/100g\;QAE$, and total flavonoid content of $3.3{\pm}0.0mg/100g\;QAE$. For the DPPH radical scavenging ability of the fermentation product also increased in a concentration dependent manner, with ORAC activity of the product showed a high activity of $20.7{\pm}0.41{\mu}M$ TE. Therefore, fermentation process can be the one of method for improving the product. The efficiency of lotus flower vinegar fermentation can be reached with an initial sugar condition of 25% (sample B).