Structural analysis of macromolecular levan produced by Bacillus megaterium GJT321 based on enzymatic method

Yu, Xiaomeng; Li, Liyan; Zhang, Jingliang; Shen, Zhaopeng; Zhu, Chunshen; Wang, Peng; Jiang, Xiaolu

doi:10.1016/j.ijbiomac.2016.09.086

Cited by 27 publications

(20 citation statements)

References 38 publications

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“…A broad band at 1642.41 cm -1 was due to the bound water [54]. C-O-H stretching vibration appears at 1123.31 cm -1 , and the strong broad peak at 1011.77 cm −1 was characterized by glycosidic linkage (C-O-C) stretching vibration in pyranose or furanose which is the carbohydrate fingerprint [55,56]. The broad peak at 924.03 and 807.27 cm -1 is confirmed as the furanose of the sugar units [57].…”

Section: Fourier-transform Infrared Spectroscopy (Ftir)mentioning

confidence: 99%

Structural Characterization of Functional Ingredient Levan Synthesized by Bacillus siamensis Isolated from Traditional Fermented Food in Thailand

Thakham

Thaweesak

Teerakulkittipong

et al. 2020

International Journal of Food Science

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The rising global population continues to threaten the world’s food security. The discovery of new technologies to produce food of nutritional and functional properties is urgently needed. One beneficial food to humans of known nutritional value is the prebiotic levan. To address the problem, the present work is aimed at isolating levansucrase enzyme-producing microorganisms from traditional fermented food in Thailand. Bacterial colony morphology was observed for mucoidal consistency on culture plates. Isolated colonies were characterized morphologically by gram staining methods. Dinitrosalicylic acid (DNS) and thin-layer chromatography (TLC) reported the highest microbial enzyme activity of 8.51 IU/ml at 12 hours via hydrolysis and frutotransferase activities. Structural characterization of levan via Fourier-transform infrared spectroscopy (FTIR) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy showed β-(2,6)-fructofuranose linkages. The highest enzyme activity was exhibited by bacterium B-6 identified as Bacillus siamensis NR 11274.1 based on the 16s rDNA gene sequence analyses. Thus, the isolated bacterium from the traditional food was confirmed to produce levansucrase enzyme of high industrial importance for the synthesis of levan as a functional food.

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Section: Fourier-transform Infrared Spectroscopy (Ftir)mentioning

confidence: 99%

Structural Characterization of Functional Ingredient Levan Synthesized by Bacillus siamensis Isolated from Traditional Fermented Food in Thailand

Thakham

Thaweesak

Teerakulkittipong

et al. 2020

International Journal of Food Science

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“…The wide and strong peak at 3304 cm − 1 was caused by the stretching vibration of O-H (Moussa et al 2017;Zhang et al 2014), indicating the existence of intermolecular hydrogen bonding. The weak peaks at 2931 cm − 1 and 2887 cm − 1 were the results of C-H stretching vibration and bending vibration respectively (Yu et al 2016). The strong absorption peak at 1644 cm − 1 was caused by the O-H bending vibration, which might be caused by the presence of water in the sample (Hao et al 2016).…”

Section: Ft-ir Analysismentioning

confidence: 93%

“…9B), seven major proton signals were observed at 4.10 ppm (H3), 4.01 ppm (H4), 3.87 ppm (H5), 3.82 ppm (H6a), 3.68 ppm (H1a), 3.60 ppm (H1b), and 3.48 ppm (H6b), respectively. All of them were in the ring proton region (3.4-4.2 ppm) indicating that the polysaccharide was composed of ketose (Cai et al 2019;Yu et al 2016). The ratio of peak areas was approximately 1:1:1:1:1:1:1 which meant the same amount of every kind of H atom.…”

Section: Nmr Analysismentioning

confidence: 95%

Biosynthesis and Structural Characterization of Levan by a Recombinant Levansucrase From Bacillus Subtilis ZW019

Wang

Zhao

et al. 2021

Preprint

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ObjectivesThe yield of levan extracted from microbial fermentation broth is low, so in vitro catalytic synthesis of levan by levansucrase is expected to be one of the industrial production approaches of levan. ResultsA recombinant plasmid Pet-28A-AcmA-Z constructed in the previous study was used to produce levansucrase. The recombinant levansucrase could be easily purified in one step and the purified enzyme had a single band clearly visible in SDS-PAGE. The conditions for enzymatic reactions was optimal at pH 5.2 and 40 ℃, and the activity of enzymes was stimulated by K+ and Ca2+. The yield of levan biosynthesis from 10% (w/v) sucrose with 6.45 U/g sucrose of levansucrase was 30.6 g/L. The molecular weight of the levan was about 1.56×106 Da, as measured by GPC. HPIC analysis showed that the monosaccharide composition of the levan was fructose and glucose. The results of FTIR and NMR analysis indicated that the polymer produced by the recombinant levansucrase was β-(2, 6) levan.ConclusionsThe results of this study provide a basis for large-scale production of levan by enzymatic method.

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“…The sample was mixed with KBr (#8966 KBR PREM SPCTROGRD FTIR PWD, Thermo Fisher Scientific) in a ratio of about 1:100, ground thoroughly and then pressed into a 1 mm pellet. The pellet was scanned 32 times in the absorbance mode from 4000 to 400 cm −1 with a 4 cm −1 resolution 6,22 …”

Section: Methodsmentioning

confidence: 99%

“…The MWs of the purified levan (redissolved with deionized water to 1 mg mL –1 ) were determined with a gel permeation chromatography (GPC) system, 3,22 which employed a Bio‐Sep 6FF Sepharose CL gel filtration column eluted with 0.7% (wt/vol) Na 2 SO 4 at a flow rate of 0.5 mL min –1 . The sugar contained in the elution fractions was determined by the phenol–sulfuric acid method 24 .…”

Section: Methodsmentioning

confidence: 99%

Factors affecting the production and molecular weight of levan in enzymatic synthesis by recombinant Bacillus subtilis levansucrase SacB‐T305A

Hou

Huang

Huí

et al. 2020

Polymer International

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Levansucrase (EC 2.4.1.10) catalyzes both levan polymerization and sucrose hydrolysis. Levan production by recombinant Bacillus subtilis levansucrase SacB‐T305A was investigated. The maximum conversion yield (33.09%) of sucrose to levan and levan yield (115.8 g L–1) were obtained as 30% (wt/vol) and 40% (wt/vol) initial sucrose, respectively. Under highly acidic conditions (pH 3.5 and below), the polymerization not only shifted distinctly to high molecular weight levan but also enlarged the size of low molecular weight levan. In particular, the proportion of high molecular weight levan to total products increased to 9‐fold at pH 3.5. The presence of Fe2+, Mg2+, Mn2+ and Zn2+ ions at common ion strength (5 mmol L–1) increased only the size of low molecular weight levan from about 20 kDa to about 35 kDa. Our findings provide a supplementary view about the mechanism of levan molecular weight distribution in enzymatic synthesis and offer a possibility for producing levan of different molecular weights by modulating enzymatic reaction conditions. © 2020 Society of Industrial Chemistry

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Structural analysis of macromolecular levan produced by Bacillus megaterium GJT321 based on enzymatic method

Cited by 27 publications

References 38 publications

Structural Characterization of Functional Ingredient Levan Synthesized by Bacillus siamensis Isolated from Traditional Fermented Food in Thailand

Structural Characterization of Functional Ingredient Levan Synthesized by Bacillus siamensis Isolated from Traditional Fermented Food in Thailand

Biosynthesis and Structural Characterization of Levan by a Recombinant Levansucrase From Bacillus Subtilis ZW019

Factors affecting the production and molecular weight of levan in enzymatic synthesis by recombinant Bacillus subtilis levansucrase SacB‐T305A

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