- Inulinase immobilisation in PAA/PEG composite for efficient fructooligosaccharides production
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Inulinase was immobilised by entrapment method in polyacrylamide/polyethylene glycol composite and evaluated for its efficiency for short-chain fructooligosaccharides (3–6 degrees of polymerisation) production in batch hydrolysis system. Aqueous two-phase
- Dimitrovski, Darko,Krastanov, Albert,Temkov, Mishela,Velickova, Elena
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- Enhancing fructooligosaccharides production by genetic improvement of the industrial fungus Aspergillus niger ATCC 20611
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Aspergillus niger ATCC20611 is one of the most potent filamentous fungi used commercially for production of fructooligosaccharides (FOS), which are prospective components of functional food by stimulating probiotic bacteria in the human gut. However, current strategies for improving FOS yield still rely on production process development. The genetic engineering approach hasn't been applied in industrial strains to increase FOS production level. Here, an optimized polyethylene glycol (PEG)-mediated protoplast transformation system was established in A. niger ATCC 20611 and used for further strain improvement. The pyrithiamine resistance gene (ptrA) was selected as a dominant marker and protoplasts were prepared with high concentration (up to 108?g?1 wet weight mycelium) by using mixed cell wall-lysing enzymes. The transformation frequency with ptrA can reach 30–50 transformants per μg of DNA. In addition, the efficiency of co-transformation with the EGFP reporter gene (egfp) was high (approx. 82%). Furthermore, an activity-improved variant of β-fructofuranosidase, FopA(A178P), was successfully overexpressed in A. niger ATCC 20611 by using the transformation system. The transformant, CM6, exhibited a 58% increase in specific β-fructofuranosidase activity (up to 507?U/g), compared to the parental strain (320?U/g), and effectively reduced the time needed for completion of FOS synthesis. These results illustrate the feasibility of strain improvement through genetic engineering for further enhancement of FOS production level.
- Zhang, Jing,Liu, Caixia,Xie, Yijia,Li, Ning,Ning, Zhanguo,Du, Na,Huang, Xirong,Zhong, Yaohua
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- A high-speed countercurrent chromatography for preparing high-purity kestose chitosan monomer
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The invention discloses a method for preparing high-purity sucrose fructan monomers by high-speed counter-current chromatography, which comprises the following steps: protecting hydroxyl of fructooligosaccharide to obtain fructooligosaccharide derivatives, carrying out high-speed counter-current chromatography to separate the fructooligosaccharide derivatives to obtain a kestose derivative, a nystose derivative and a glucopyranoside derivative, removing the hydroxyl protecting group to obtain the high-purity kestose, nystose and glucopyranoside. The purities of the kestose, nystose and glucopyranoside are respectively higher than 98%. The method has the advantages of short separation time, high preparation quantity,, high recovery rate, no sample loss, mild separation environment and solvent saving, and can implement scale-up production.
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Paragraph 0038-0042
(2017/09/26)
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- Optimization of levansucrase/endo-inulinase bi-enzymatic system for the production of fructooligosaccharides and oligolevans from sucrose
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A bi-enzymatic system based on the combined use of levansucrase (LS) from Bacillus amyloliquefaciens and endo-inulinase from Aspergillus niger in a one-step reaction was investigated for the synthesis of fructooligosaccharides (FOSs) and oligolevans using sucrose as the sole substrate. Sucrose concentration was the most important independent variable, whilst LS to endo-inulinase ratio exhibited significant effects on the end-product profiles. The interaction between sucrose concentration and reaction time exhibited significant effect on all responses. At the initial stage of time course, short chain FOSs (scFOSs, 1-kestose, nystose, 1F-fructosylnystose) were the major products, whilst 6-kestose, medium chain fructooligosaccharides (mcFOSs, levanohexaose, levanopentaose) and oligolevans became the dominant ones at the late stage. The optimal conditions leading to a high yield of scFOSs (1:1 ratio, 0.5 h, 0.6 M) were different from those resulting in a high yield of mcFOSs and oligolevans (1.85:1 ratio, 1.77 h, 0.6 M). The bi-enzymatic system has a great potential for the production of FOSs and oligolevans at a large scale because of its high yield (57-65%, w/w) and productivity (65.8-266.8 g/L h), and its uses of low temperature (35 °C) and low concentration of sucrose. To the best of our knowledge, this is the first study on the optimization of a LS/endo-inulinase bi-enzymatic system.
- Tian, Feng,Khodadadi, Maryam,Karboune, Salwa
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- Screening of biocatalysts for transformation of sucrose to fructooligosaccharides
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Twenty microorganisms comprising of sixteen molds, two yeasts and two bacteria were evaluated for their ability to produce fructosyltransferase (FTase) and generate fructooligosaccharides (FOS) from sucrose. FTase production by these microorganisms was studied over a period of 120 h on medium containing 20% (w/v) sucrose as the sole carbon source. High FTase levels (35-31 U/ml) were observed in culture filtrates of Aspergillus flavus, Aspergillus niger, Aspergillus terreus and Penicillium islandicum. Higher concentrations of FOS were generated from 50% (w/v) sucrose using culture filtrates of A. flavus NFCCI 2364 (63.40%, w/w), A. niger SI 19 (54.94%, w/w), A. flavus NFCCI 2785 (44.61%, w/w), P. islandicum MTCC 4926 (43.56%, w/w), A. terreus NFCCI 2347 (24.17%, w/w) and Fusarium solani NFCCI 2315 (15.25%, w/w). Kestose, nystose and 1-fructofuranosyl nystose were the predominant oligosaccharides as revealed by HPLC analysis.
- Ganaie, Mohd Anis,Gupta, Uma Shanker,Kango, Naveen
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- Potential application of commercial enzyme preparations for industrial production of short-chain fructooligosaccharides
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Twenty-five commercial enzyme preparations for use in the food industry were assayed for transfructosylation activity. Three preparations showed high transfructosylation activity from sucrose as well as the ratio of transferase and hydrolase activities. Short-chain fructooligosaccharides (sc-FOS) were not hydrolyzed by the three enzyme preparations after a 12 h reaction time. At a 6 h reaction time, yield and volumetric productivity were in the range from 58.8 to 62.6% (g sc-FOS/100 g initial sucrose) and 52.5 to 55.9 g sc-FOS/L h, respectively. One enzyme preparation was then evaluated for sc-FOS synthesis. Thus, environmental factors influencing the reaction were studied on products. Total sc-FOS concentration was not affected by temperature, pH and enzyme concentration at the studied levels, but high concentrations of sucrose affected the sc-FOS formation. The results suggest that these enzyme preparations can be exploited as a source of food-grade fructosyltransferase, in addition to Pectinex Ultra SP-L.
- Vega-Paulino,Zuniga-Hansen
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experimental part
p. 44 - 51
(2012/04/11)
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- PROCESS OF CITRIC OR PHOSPHORIC PARTIAL HYDROLYSIS OF INULIN FOR THE OBTENTION OF FRUCTOOLIGOSACCHARIDES - FOS
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In the technical field of Chemistry, encompassing products defined like FOS - fructooligosaccharides and applicable in food and or medical-pharmaceutical, veterinary and odontological industries, including the segment of supplements and functional foods, through the partial hydrolysis of inulins and similar fuctans till hydrossoluble FOS in the preferential range of DP - Degree of Polymerization from 2 to 18 or even more with reduced content of free fructose, using as catalysts for hydrolysis the citric and / or phosphoric acids as alternative to the classic hydrochloric or sulfuric acids and to the microbial enzymes, with the particularity that, through partial neutralization or simple dilution of the hydrolyzates, the catalysts now proposed, may either remain for the industrial and ulterior proposals or alternatively be removed by ion exchange resins or other means, being still possible that the citric or phosphoric acid may be conveniently diluted to the preferential range of pH 2.0 to 3.0 as moderate acidicity and the hydrolysis be carried out at moderate temperatures in the range from 75oC to 90oC in the range of preferential times from 5 to 30 minutes.
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Page/Page column 25-27
(2009/03/07)
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- Physicochemical characterization of fructooligosaccharides and evaluation of their suitability as a potential sweetener for diabetics
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Fructooligosaccharides (FOSs) were prepared from sucrose using fungal fructosyl transferase (FTase) obtained from Aspergillus oryzae MTCC 5154. The resulting mixture consisted of glucose (28-30%), sucrose (18-20%) and fructooligosaccharides (50-54%) as indicated by HPLC analysis. Identification of oligomers present in the mixture of fructooligosaccharides was carried out using NMR spectroscopy and LC-MS. No compounds other than mono-, di-, tri-, tetra- and pentasaccharides were identified in the FOS mixture prepared using FTase. NMR and LC-MS spectra proved the absence of any toxic microbial metabolites of Aspergillus species in FOS thereby emphasizing its safe use as a food ingredient. Animal studies conducted on streptozotocin-induced diabetic rats suggested that the use of FOS as an alternative non-nutrient sweetener is without any adverse effects on various diabetes-related metabolic parameters. Despite the high free-sugar content associated with it, FOS did not further aggravate the hyperglycemia and glucosuria in diabetic animals, even at 10% levels. On the other hand, by virtue of its soluble fibre effect, it has even alleviated diabetic-related metabolic complications to a certain degree.
- Mabel,Sangeetha,Platel, Kalpana,Srinivasan,Prapulla
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