• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1880-1893
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• 2015

In this study, Pseudomonas R0-14, which was isolated from Arctic soil samples, showed a clear halo when grown on M9 medium agarose plates containing olive oil-rhodamine B as substrate, suggesting that it expressed putative lipase(s). A putative lipase gene, lipR, was cloned from R0-14 by genome walking and Touchdown PCR. lipR encodes a 562-amino-acid polypeptide showing a typical α/β hydrolase structure with a catalytic triad consisting of Ser₁₅₃-Asp₂₀₂-His₂₆₀ and one α-helical lid (residues 103-113). A phylogenetic analysis revealed that LipR belongs to the lipase subfamily I.3. LipR was successfully expressed in Escherichia coli, purified, and biochemically characterized. Recombinant LipR exhibited its maximum activity towards p-nitrophenyl butyrate at pH 8.5 and 60℃ with a K_m of 0.37 mM and a k_cat of 6.42 s^-1. It retained over 90% of its original activity after incubation at 50℃ for 12 h. In addition, LipR was activated by Ca²⁺, Mg²⁺, Ba²⁺, and Sr²⁺, while strongly inhibited by Cu²⁺, Zn²⁺, Mn²⁺, and ethylenediaminetetraacetic acid. Moreover, it showed a certain tolerance to organic solvents, including acetonitrile, isopropanol, acetone, methanol, and tert-butanol. When algal oil was hydrolyzed by LipR for 24 h, there was an enrichment of n-3 long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (1.22%, 1.65-fold), docosapentaenoic acid (21.24%, 2.04-fold), and docosahexaenoic acid (36.98%, 1.33-fold), and even a certain amount of diacylglycerols was also produced. As a result, LipR has great prospect in industrial applications, especially in food and/or cosmetics applications.

• Analytical Science and Technology
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• v.6 no.1
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• pp.9-19
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• 1993

Fatty acid salts derived from soap can be transferred into a typical derivative with p-bromophenacyl bromide using crown ether, a catalizer by the solid-liquid phase transfer reaction in nonpolar, aprotic solvents and separated by the reverse phase high performance liquid Chromatography (RP-HPLC) and determined using UV detector. The minimal limit of detection was defined at approximately 10~50ng in accordance with the chain length. The derivatization reaction in the presence of EDTA can be applied mot only to the calcium salts but also to the other various metal salts. The recoveries of fatty acid derivatizations in the absence and presence of the midium containing the yeast extract were obtained $95.4{\pm}1.2$, and $85.2{\pm}2.4%$ respectively. The analytical method would be applicable to determine the biodegradation of fatty acid salts in nature as well as in artificial condition such as shaker flask-medium method.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.252-257
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• 1994

A new hydrogen sulfide-oxidation bacterium, Thiobacillus sp. was isolated from waste coal mine water around Hawsun in Chunnam province. The isolate was motile gram-negative rod shape, formed spore and grew up to be aerobically facultative chemolithotroph by using energy released from the oxidation of reduced inorganic sulfur compounds. It could assimilate various kinds of organic compounds and grew well upon thiosulfate-supplemented basal medium. To the lelvel of 32 mM in thiosulfate concentration, thiosulfate in itself was utilized as energy source for growth. However, from those of the higher concentration than 32 mM, thiosulfate functioned specifically as the substrate inhibitor rather than as the energy source. It was found that the optimum thiosulfate concentration for growth was 32 mM. The G+C content of the DNA was 65.0 mol%. The isolate had 16 : 1 + 17$_{cyc}$, 16 : 0 as their major non-hydroxylated cellular fatty acids, 3-OH 12 : 0 as a hydroxylated fatty acid and also contained unidentified $C_{18}$ branched fatty acid. The ubiquinone system in the respiratory chain was Q-9. Based on the physiological and biochemical characteristics, the isolate was assigned to a novel species of the genus Thiobacillus sp. iw.

• Animal Bioscience
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• v.35 no.9
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• pp.1408-1417
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• 2022

Objective: The objective of this study was to determine whether dietary black soldier fly (Hermetia illucens, HI) larvae oil (HILO) could serve as an alternative fat source to soybean oil (SBO) in laying hen diets. Methods: We randomly assigned 25-week-old Hy-line Brown laying hens (n = 144) to receive (n = 6 hens/group; eight replicates) a control or an experimental diet in which SBO was replaced with 50% (50HILO) or 100% HILO (100HILO). Results: Dietary HILO did not negatively affect body weight or productive performance during the study. The eggs also had similar quality parameters, proximate composition, and cholesterol levels. However, the yolk color index was significantly higher (p<0.01) in the 100HILO than in the other groups. Dietary HILO significantly altered the composition of fatty acids (FAs) in abdominal fat and eggs. Total saturated fatty acids (SFAs) and total polyunsaturated FAs (PUFAs) were significantly increased and decreased in the 50HILO and 100HILO groups, respectively, compared with those in the control group (p<0.001 and p<0.0001, respectively). Specifically, the medium-chain FAs lauric and myristic acids were remarkably increased in the abdominal fat of laying hens fed HILO (p<0.0001), whereas only myristic acid increased in eggs (p<0.0001). Undesirable heavy metal (aluminum, fluorine, arsenic, lead, mercury, and cadmium) concentrations were below permissible limits in eggs. Conclusion: We considered that HILO could be an alternative dietary fat to SBO for laying hens with maintained productive performance and good egg quality.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1120-1124
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• 2005

Pseudomonas putida KCTC 1639 was newly identified as a potential producer of biodegradable medium chain length polyhydroxyalkanoates. It exhibited a carbon assimilation pattern quite different from other known P. putida strains, but a more similar pattern with P. oleovorans, which assimilates the carbon sources mainly through ${\beta}$-oxidation rather than the fatty acid biosynthesis pathway. The PHA synthesis gene cluster from P. putida KCTC1639 was composed of two gene loci; the PHA synthase gene locus and granule-associated gene locus, which were cloned and deposited in the GenBank under accession numbers AY286491 and AY750858 as a new nucleotide sequence, respectively. The molecular structure and amino acid homology of the new gene cluster were compared with those from Pseudomonas species, including other P. putida strains and P. oleovorans, and a higher than $90\%$ homology was observed.

• Food Science and Industry
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• v.51 no.2
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• pp.148-156
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• 2018

Structured lipids are lipids in which the composition and/or positional distribution of fatty acids have been chemically or enzymatically modified from their natural biosynthetic form. Because structured lipids have desired nutritional, physicochemical, textural or physiological properties for applications in processed foods, functional foods, or nutraceuticals, many research activities have been aimed at their commercialization. The enzymatic production of structured lipids using lipases as the biocatalysts has a big potential in the future market due to the specificity or selectivity of the lipases. This article introduced some examples of specialty structured lipids that have been enzymatically produced and have been utilized as commercialized products. The commercialized products include medium- and long-chain triacylglycerols, human milk fat substitutes, cocoa butter equivalents, trans-free plastic fats, low-calorie fats/oils, and health-beneficial fatty acid-rich oils.

• Korean Journal of Microbiology
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• v.41 no.3
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• pp.225-231
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• 2005

The characteristics of cell growth and medium-chain-length polyhydroxyalkanoate (MCL-PHA) biosynthesis of Pseudomonas chlororaphis HS21 were investigated using plant oils as the carbon substrate. The organism was efficiently capable of utilizing plant oils, such as palm oil, corn oil, and sunflower oil, as the sole carbon source for growth and MCL-PHA production. When palm oil (5 g/L) was used as the carbon source, the cell growth and MCL-PHA accumulation of this organism occurred simultaneously, and a high dry cell weight (2.4 g/L) and MCL-PHA ($40.2\;mol{\%}$ of dry cell weight) was achieved after 30 hr of batch-fermentation. The repeating unit in the MCL-PHA produced from palm oil composed of 3-hydroxyhexanoate ($7.0\;mol{\%}$), 3-hydroxyoctanoate ($45.3\;mol{\%}$), 3-hydroxydecanoate ($39.0\;mol{\%}$), 3-hydroxydodecanoate ($6.8\;mol{\%}$), and 3-hydroxytetradecanoate ($1.9\;mol{\%}$), as determined by GC/MS. Even though glucose was a carbon substrate that support cell growth but not PHA production, the conversion rate of palm oil to PHA was significantly increased when glucose was fed as a cosubstrate, suggesting that bioconversion of some functionalized carbon substrates to related polymers in P chlororaphis HS21 could be enhanced by the co-feed of good carbon substrates for cell growth. In addition, the change of compositions of repeating units in MCL-PHAs synthesized from the plant oils was markedly affected by the supplementation of acrylic acid, an inhibitor of fatty acid ${\beta}-oxidation$. The addition of acrylic acid resulted in the increase of longer chain-length repeating units, such as 3-hydroxydodecanoate and 3-hydroxytetradecanoate, in the MCL-PHAs produced. Particularly, MCI-PHAs containing high amounts of unsaturated repeating units could be produced when sunflower oil and corn oil were used as the carbon substrate. These results suggested that the alteration of PHA synthesis pathway by acrylic acid addition can offer the opportunity to design new functional MCL-PHAs and other unusual polyesters that have unique physico-chemical properties.

• Journal of the Korean Applied Science and Technology
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• v.1 no.1
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• pp.49-55
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• 1984

The lipid and fatty acid compositions and vitamin E content were analyzed in 48 human mature milk, 3 infant formulas (modified milk powder) and 8 market milk samples. The total lipid content in modified milk powder was similar those in human milk and market milk when total solids content of modified milk powder was corrected to that of human milk. In comparison with human milk, modified milk powder contained a lower proportion of triglycerides and higher proportions of phospholipids, free fatty acids and cholesterol esters. The ratios of phospholipids/triglycerides, total cholesterol / triglycerides and total tocopherol/total lipids in modified milk powder were significantly higher than those in human milk and market milk. The American recommendation for linoleic acid (0.7 IU/g) could be satisfied with human milk, modified milk powder and market milk. The proportions of short- and medium-chain even numbered saturates were higher and the proportions of long-chain derivatives of linoleic (${\omega}6$ series) and linolenic (${\omega}3$ series)acids were lower in modified milk powder and market milk than in human milk. It is concluded that in view of their levels in breast milk, the polyenoic derivatives of linoleic and linolenic acids must be taken into account when assessing infant foods.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.8
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• pp.1110-1117
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• 2001

Responses of the rumen fungus, Neocallimastix frontalis RE1, to long chain fatty acid (LCFA) were evaluated by measuring gas production, filter paper (FP) cellulose digestion and polysaccharidase enzyme activities. LCFA (stearic acid, $C_{18:0}$; oleic acid, $C_{18:1}$; linoleic acid, $C_{18:2}$ and linolenic acid, $C_{18:3}$) were emulsitied by ultrasonication under anaerobic condition, and added to the medium. When N frontalis RE1 was grown in culture with stearic, oleic and linoleic acid, the cumulative gas production, gas pool size, FP cellulose digestion and enzymes activities significantly (p<0.05) increased at some incubation times(especially, exponential phases of fungal growth, 48~120 h of incubation) relative to that for control cultures. However, the addition of linolenic acid strongly inhibited all of the investigated parameters up to 120 h incubation, but not after 168 and 216 h of incubation. These results indicated that stearic, oleic and linoleic acids tended to have great stimulatory effects on fungal cellulolysis, whereas linolenic acid caused a significant (p<0.05) inhibitory effects on the cellulolysis by the rumen fungus. These results are the first report of the effect of LCFAs on the ruminal fungi. Further research is needed to identify the mode of action of LCFAs on fungal strains and to verify whether or not ruminal fungi have ability to hydrate unsaturated LCFAs to saturated FAs. There was high correlation between cumulative in vitro gas production and fungal growth (94.78%), FP cellulose degradation (96.34%), CMCase activity(90.86%) or xylanase activity (87.67%). Thus measuring of cumulative gas production could be a useful tool for evaluating fungal growth and/or enzyme production by ruminal fungi.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.995-1002
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• 2000

To evaluate energy efficiency and partition of nutrients, 32 piglets were weaned at 14 d of age and individually fed diets containing 15% fat from coconut oil (CO, medium chain saturated), high oleate sunflower oil (HOSO, n-9 series), soybean oil (SO, n-6 series), or linseed oil plus fish oil, (LF, n-3 series). After 4 weeks, the subjects were sacrificed to evaluate empty body composition and apparent ileal digestibility with the slaughter method. No statistical effect of dietary fat sources on growth was observed. The digestibility of fat from the coconut oil diet was higher than fats from the diets containing high levels of unsaturated fatty acids. The efficiency of use of metabolizable energy for growth averaged 63% and was not affected by the diet. Dietary fat composition was reflected strongly in backfat. Total body neutral and polar fatty acids were influenced too. For the whole body phospholipid fraction the ratio of n-6 to n-3 and the double bond index were 4.3, 5.8, 7.2, 0.78 and 69, 87, 89, 87 for CO, HOSO, SO, and LF respectively. These results show that for the coconut oil diet the degree of unsaturation of phospholipids in the body was lower and that, in the other diets, it did not differ, but double bond index was maintained with different n-6 to n-3 ratios in carcass fat. On the whole the data on body fat composition indicate that the dietary fat tended to be deposited in similar quantity in the body, whatever was the dietary fatty acid profile.