Enzymatic analysis of levan produced by lactic acid bacteria in fermented doughs.

Levans and inulins are fructans with mainly ß-(2→6) and ß-(2→1) linkages, respectively. Levans are produced by many lactic acid bacteria, e.g. during sourdough fermentation. Levans have shown prebiotic properties and may also function as in situ-produced hydrocolloids. So far, levan contents have been measured by acid hydrolysis, which cannot distinguish levans from e.g. inulins. In order to develop a specific analysis for levan in food matrices, a Paenibacillus amylolyticus endolevanase was combined with exoinulinase for levan hydrolysis. A separate endoinulinase treatment was used to detect the possible presence of inulin. Interfering sugars were removed by a pre-wash with aqueous ethanol. Levan content was estimated from fructose and glucose released in the hydrolysis, with a correction made for the residual fructose and glucose-containing sugars. The method was validated using wheat model doughs spiked with commercial Erwinia levan, and tested by analyzing levan content in Leuconostoc mesenteroides DSM 20343-fermented fava bean doughs.

Optimization of Isomaltooligosaccharide Size Distribution by Acceptor Reaction of Weissella confusa Dextransucrase and Characterization of Novel α-(1→2)-Branched Isomaltooligosaccharides.

Long-chain isomaltooligosaccharides (IMOs) are promising prebiotics. IMOs were produced by a Weissella confusa dextransucrase via maltose acceptor reaction. The inputs of substrates (i.e., sucrose and maltose, 0.15-1 M) and dextransucrase (1-10 U/g sucrose) were used to control IMO yield and profile. According to response surface modeling, 1 M sucrose and 0.5 M maltose were optimal for the synthesis of longer IMOs, whereas the dextransucrase dosage showed no significant effect. In addition to the principal linear IMOs, a homologous series of minor IMOs were also produced from maltose. As identified by MS(n) and NMR spectroscopy, the minor trisaccharide contained an α-(1→2)-linked glucosyl residue on the reducing residue of maltose and thus was α-d-glucopyranosyl-(1→2)-[α-d-glucopyranosyl-(1→4)]-d-glucopyranose (centose). The higher members of the series were probably formed by the attachment of a single unit branch to linear IMOs. This is the first report of such α-(1→2)-branched IMOs produced from maltose by a dextransucrase.

Lactose- and cellobiose-derived branched trisaccharides and a sucrose-containing trisaccharide produced by acceptor reactions of Weissella confusa dextransucrase.

Dextran-producing Weissella have received significant attention. However, except for maltose, the acceptor reactions of Weissella dextransucrases with different sugars have not been investigated. The action of recombinant Weissella confusa VTT E-90392 dextransucrase was tested with several potential acceptors, particularly, analogs lactose and cellobiose. The major acceptor products of both disaccharides were identified as branched trisaccharides, with a glucosyl residue α-(1 → 2)-linked to the acceptor's reducing end. An additional product, isomelezitose (6(Fru)-α-Glcp-sucrose), was also produced when using lactose as an acceptor. This is the first report of the synthesis of isomelezitose by a dextransucrase. The NMR spectra of the three trisaccharides were fully assigned, and their structures were confirmed by selective enzymatic hydrolysis. The trisaccharides prepared from (13)C6(glc) sucrose and lactose were analyzed by ESI-MS(n), and the fragmentation patterns of these compounds were characterized.

Cloning and characterization of a Weissella confusa dextransucrase and its application in high fibre baking.

Wheat bran offers health benefits as a baking ingredient, but is detrimental to bread textural quality. Dextran production by microbial fermentation improves sourdough bread volume and freshness, but extensive acid production during fermentation may negate this effect. Enzymatic production of dextran in wheat bran was tested to determine if dextran-containing bran could be used in baking without disrupting bread texture. The Weissella confusa VTT E-90392 dextransucrase gene was sequenced and His-tagged dextransucrase Wc392-rDSR was produced in Lactococcus lactis. Purified enzyme was characterized using (14)C-sucrose radioisotope and reducing value-based assays, the former yielding K(m) and V(max) values of 14.7 mM and 8.2 µmol/(mg ∙ min), respectively, at the pH optimum of 5.4. The structure and size of in vitro dextran product was similar to dextran produced in vivo. Dextran (8.1% dry weight) was produced in wheat bran in 6 h using Wc392-rDSR. Bran with and without dextran was used in wheat baking at 20% supplementation level. Dextran presence improved bread softness and neutralized bran-induced volume loss, clearly demonstrating the potential of using dextransucrases in bran bioprocessing for use in baking.

Reduction of diazinon and dimethoate in apple juice by pulsed electric field treatment.

BACKGROUND: Organophosphorus pesticides (OPPs) are widely used in agricultural production in China, and residues of OPPs in agro-products and foods have become a public health concern. Chronic exposure to OPPs can result in potential immunosuppressive effects, cytotoxicity and mutagenicity. Pulsed electric fields (PEFs) have the potential to be used as an alternative to conventional techniques of food production. The aim of the present study was to investigate the influence of PEFs on the degradation of diazinon and dimethoate added to apple juice. RESULTS: PEF treatment significantly promoted the degradation of both pesticides (P < 0.05). The extent of degradation was strongly influenced by the electric field strength and treatment time, and maximum degradation of both diazinon (47.6%) and dimethoate (34.7%) was achieved by treatment at 20 kV cm⁻¹ for 260 µs. The degradation behaviour of the pesticides was described by Weibull and Hülsheger models, both of which successfully fitted the degradation of diazinon and dimethoate. In addition, the decline in the toxicity of samples was observed with a photobacterium bioassay. CONCLUSION: PEF treatment was effective in eliminating residues of diazinon and dimethoate spiked in apple juice and in mitigating sample toxicity.

The degradation of chlorpyrifos and diazinon in aqueous solution by ultrasonic irradiation: effect of parameters and degradation pathway.

In this paper, elimination of two types of organophosphorus pesticides (OPPs), chlorpyrifos and diazinon spiked in aqueous solution by ultrasonic irradiation was investigated. Results showed that chlorpyrifos and diazinon could be effectively and rapidly degraded by ultrasonic irradiation, and the degradation of both pesticides was strongly influenced by ultrasonic power, temperature and pH value. Furthermore, two and seven products for the degradation of chlorpyrifos and diazinon formed during ultrasonic irradiation have been identified by gas chromatography-mass spectrometry, respectively. The hydrolysis, oxidation, hydroxylation, dehydration and decarboxylation were deduced to contribute to the degradation reaction and the degradation pathway for each pesticide under ultrasonic irradiation was proposed. Finally, the toxicity evaluation indicated that the toxicity decreased for diazinon solution after ultrasonic irradiation, but it increased for chlorpyrifos solution. The detoxification of OPPs by ultrasonic irradiation was discriminative.

Degradation of diazinon in apple juice by ultrasonic treatment.

The degradation of diazinon spiked in apple juice treated by ultrasonic treatment was investigated in this paper. Results showed that the ultrasonic power and initial concentration of diazinon significantly influenced the degradation percentage of diazinon (p<0.05) and the degradation of diazinon followed the first-order kinetics model well. Based on the gas chromatography-mass spectrometry (GC-MS) analysis, seven degradation products of diazinon have been identified. A degradation pathway involving hydrolysis of the ester moiety, oxidation, hydroxylation, dehydration, and decarboxylation was proposed. Simultaneously, the toxicity of apple juice was mitigated by the ultrasonic treatment based on the photobacterium bioassay.