Novel bioconversion of sodium glutamate to γ-poly-glutamic acid and γ-amino butyric acid in a mixed fermentation using Bacillus subtilis HA and Lactobacillus plantarum K154

Both γ-poly-glutamic acid (γ-PGA) and γ-amino butyric acid (GABA) were produced from sodium glutamate in a mixed fermentation. A culture broth was obtained using a defined medium including 3% glutamate and Bacillus subtilis HA for 3 days at 42°C, with 1.20×10⁸ CFU/mL viable cells, a 0.46 Pa·sⁿ consistency index, and a 1.94% mucilage content. In a second fermentation using Lactobacillus plantarum K154 at 37°C for 3 days, 1.29% of the remaining glutamic acid in the viscous culture broth was converted to 0.86% GABA. Glutamic acid in the first culture broth fortified with 5% skim milk was completely converted to GABA. The final co-cultured broth had 0.48% GABA, a 0.294% tyrosine content, and viable cells of 1.06×10⁹ CFU/mL (L. plantarum K154) and 5.42×10⁵ CFU/mL (B. subtilis HA). Serial co-culturing of these two bacteria can provide novel ingredients fortified with γ-PGA, GABA, peptides, and probiotics.     

Cloning and characterization of the Hansenula polymorpha homologue of the Saccharomyces cerevisiae PMR1 gene

A gene homologous to Saccharomyces cerevisiae PMR1 has been cloned in the methylotrophic yeast Hansenula polymorpha. The partial DNA fragment of the H. polymorpha homologue was initially obtained by a polymerase chain reaction and used to isolate the entire gene which encodes a protein of 918 amino acids. The putative gene product contains all ten of the conserved regions observed in P-type ATPases. The cloned gene product exhibits 60·3% amino acid identity to the S. cerevisiae PMR1 gene product and complemented the growth defect of a S. cerevisiae pmr1 null mutant in the EGTA-containing medium. The results demonstrate that the H. polymorpha gene encodes the functional homologue of the S. cerevisiae PMR1 gene product, a P-type Ca²⁺-ATPase. The DNA sequence of the H. polymorpha homologue has been submitted to GenBank with the Accession Number U92083. © 1998 John Wiley & Sons, Ltd.     

Inactivation of Listeria monocytogenes on frankfurters by monocaprylin alone or in combination with acetic acid

The antilisterial activity of monocaprylin (MC) and its combination with acetic acid (AA) on frankfurters was investigated. Each frankfurter was surface inoculated with a three-strain mixture of Listeria monocytogenes to obtain an inoculation level of 4.0 log CFU per frankfurter, and then dipped for 35 s in sterile deionized water (45 or 50 degrees C) containing 1% ethanol (control), 50 mM MC plus 1% ethanol, 1% AA plus 1% ethanol, or 50 mM MC plus 1% AA plus 1% ethanol. Samples were vacuum packaged, stored at 4 degrees C for 77 days, and analyzed for L. monocytogenes. Sensory odor and color of frankfurters were evaluated using a 9-point hedonic scale. Color was also objectively measured using the Minolta Chroma Meter. From day 0 to day 77, population counts of L. monocytogenes on frankfurters dipped in antimicrobial solutions at 50 degrees C were consistently lower than the control counts. Similar results were observed for samples treated at 45 degrees C. However, L. monocytogenes grew readily on control samples at both temperatures. Dipping of frankfurters in antimicrobial solutions (45 or 50 degrees C) significantly reduced (P < 0.05) the populations of L. monocytogenes. After 70 days of storage, L. monocytogenes was completely killed in samples dipped in MC+AA solution at 50 degrees C. The antimicrobial treatments did not affect the odor or color of the samples (P > 0.05). Overall, results indicated that dipping of frankfurters with MC reduced L. monocytogenes, and inclusion of AA further enhanced MC antilisterial activity, without any negative effect on odor or color.     

Production and Characterization of Branched Oligosaccharides from Liquefied Starch by the Action of B. Licheniformis Amylase

A branched oligosaccharides (BOS) mixture was produced from liquefied starch solution using a maltogenic amylase of Bacillus licheniformis (BLMA). The BOS mixture was produced by both α-1,4-bond hydrolyzing and α-1,6-transglycosylation activities of BLMA, and it contained 58.3% of various branched oligosaccharides. Small branched oligosaccharides such as isomaltose, isopanose, and panose were identified in the mixture by various analyses including high performance ion-chromatography (HPIC). Major branched DP4 and DP5 molecules in the mixture were determined as 6²-O-α-maltosylmaltose, 6³-O-α-maltosyl-maltotriose and 6²-O-α-maltotriosyl-maltose, respectively. Time course study of BOS production suggested that the hydrolysis and transglycosylation reactions catalyzed by BLMA were coupled. BLMA was likely to transfer a sugar moiety hydrolyzed from a non-reducing end of maltooligosaccharide, mainly maltose, to another moiety of sugar via the formation of α-1,6-linkage. Immobilization of BLMA was attempted as an effort to achieve a continuous process for BOS production. the immobilized enzyme showed improved thermal stability and slight loss of enzyme activity was observed during repeated usage.     

HPLC-ELSD analysis of 18 platycosides from balloon flower roots (Platycodi Radix) sourced from various regions in Korea and geographical clustering of the cultivation areas

An effective HPLC method to analyse platycosides from the balloon flower root was developed using ELSD. The optimum resolution of the platycosides was achieved on an ODS column with gradient elution of eluent A, 30 mM ammonium acetate buffer (pH 4.81): methanol: acetonitrile = 75:5:20 (v/v/v), and B, 69:5:26 (v/v/v). Amongst 18 platycosides, platycoside E showed the highest content, followed by polygalacin D₂ and 3″-O-acetylplatyconic acid A. The sum of these three compounds was recommended for quality control of balloon flower root for medicinal purposes. The samples could be clustered into groups based on platycoside content. Group I, characterised by a high concentration of platycosides, was located near the west coast of Korea, whereas group II, characterised by a low concentration of platycosides, was located inland or in mountainous area. The method could be used to control the quality of balloon flower root.     

Catalytic properties of a lipase from Photobacterium lipolyticum for biodiesel production containing a high methanol concentration

Biodiesel, an alternative fuel, is generated via the transesterification reaction of vegetable oil or animal oil with alcohol. Currently, many reports have noted that microbial lipases might be utilized for the production of biodiesel. Among them, immobilized Candida antarctica lipase B (Novozym435) is frequently utilized for its biocatalytic efficiency and availability. However, as the enzyme is unstable in a medium containing high concentrations of methanol, a multi-stepwise methanol supply is required for the efficient production of biodiesel. Photobacterium lipolyticum lipase (M37) was determined to be quite stable in a medium containing a high concentration of methanol. The enzyme activity was maintained for longer than 48 h without any loss at a methanol concentration of 10%. In an effort to evaluate enzyme performance in the production of biodiesel, we have compared M37 lipase and Novozym435 in the biodiesel production reaction using fresh or waste oil and methanol. In the 3-stepwise methanol feeding method generally conducted for Novozym435 in biodiesel production, the M37 lipase showed a similar or superior conversion yield to Novozym435. However, the M37 lipase evidenced significantly higher conversion yields in the 2 and 1 step methanol feeding reactions. Particularly in the 1 step process using 10% of methanol where almost no conversion was detected by Novozym435, the biodiesel yield achieved with M37 lipase reached a level of up to 70% of the possible maximum yield. Consequently, this methanol-tolerant lipase, M37, has been shown to be a suitable enzyme for use in the biodiesel production process.     

A comparative study on the antioxidative and anti‐allergic activities of fresh and aged black garlic extracts

The antioxidative and anti‐allergic activities of fresh and aged black garlic extracts were investigated. The garlic samples were extracted with 70% ethanol (v/v) and the total phenolic content was measured. The antioxidant capacity of extracts was assessed by determining the scavenging activities on 1,1‐diphenyl‐2‐picrylhydrazyl and hydroxyl radicals, ferricyanide reducing power, ferrous ion‐chelating ability and inhibitory effect on linoleic acid peroxidation. The anti‐allergic activity of extracts was analysed by measuring their inhibitory effects against β‐hexosaminidase release. The aged black garlic exhibited significantly higher phenolic content and greater antioxidative activity than fresh garlic. Both garlic extracts showed strong antioxidant capacity in a dose‐dependent manner. On the other hand, a considerably higher suppression of β‐hexosaminidase release was found in fresh garlic extract at lower concentration compared with that of the black garlic. Results of this study illustrate that ageing of garlic could enhance its antioxidant capacity, but could decrease its anti‐allergic activity.     

Antioxidative activities of bran extracts from twenty one pigmented rice cultivars

Ethanol–water (70:30, v/v) extracts from the bran of rice seeds from twenty one pigmented and one nonpigmented rice cultivars were evaluated for antioxidative activities using the following tests: inhibition of peroxidation of linoleic acid; inhibition of peroxidation of rabbit lipid erythrocyte membranes; reduction of potassium ferricyanide, and scavenging of superoxide anions and hydroxyl radicals. With some exceptions, extracts from the pigmented rice seeds had higher antioxidative activity than did the nonpigmented variety. The following pigmented cultivars had the highest antioxidative activities in all tests: Jumlalocal-1, Parnkhari 203, DZ78, LK1-3-6-12-1-1, and Elwee. A significant correlation was also noted between reducing power, inhibition of erythrocyte ghost membrane peroxidation, and superoxide anion and hydroxyl radical scavenging. The results suggest that: (a) ferricyanide reducing power might be a useful and simple index for large-scale evaluation of antioxidative potencies of natural products present in rice; (b) pigmented rice varieties with high antioxidative activities provide a source of antioxidants and a genetic resource to develop new health-promoting rice cultivars.