Low Digestion Property of Amylosucrase-modified Waxy Adlay Starch

김은정,김하람,최승준,박천석,문태화 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.2

Structural and digestion properties of amylosucrase-modified waxy adlay starch were investigated. The unique reaction of amylosucrase caused a decrease and an increase in the proportion of short chains and long chains, respectively, via attachment of glucosyl units to the non-reducing ends of branch chains. The in vitro digestion profile of amylosucrase-modified starch revealed that elongated branch chains were the main reason for high contents of slowly digestible and resistant starches due to formation of a more perfect crystalline structure via easy association between elongated branch chains. The glucose response in mice after consumption of amylosucrase-modified starch was similar to the response for commercial resistant starch with a gradual increase followed by a gradual decrease in blood glucose concentrations over a prolonged time. Both in vitro and in vivo tests were used to verify increased resistance to digestive enzymes caused by amylosucrase modification.

Kinetic studies of <i>in vitro</i> digestion of amylosucrase-modified waxy corn starches based on branch chain length distributions

Kim, Ha Ram,Choi, Seung Jun,Park, Cheon-Seok,Moon, Tae Wha Elsevier 2017 Food hydrocolloids Vol.65 No.-

<P><B>Abstract</B></P> <P>Classification of starch into rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS) has been in controversy. Therefore, this study aimed to ascertain the individual existence of RDS and SDS using amylosucrase-modified starch. Enzymatic hydrolysis curves of modified waxy corn starches were obtained and fitted to a logarithm of slope (LOS) plot. LOS plots for amylosucrase-modified starches revealed a discontinuity, reflecting the change of rate constant <I>k</I> during digestion, such that distinguishable <I>k</I> s for each of the phases were defined (<I>k</I> <SUB> <I>RDS</I> </SUB>, <I>k</I> <SUB> <I>SDS</I> </SUB>). The LOS plot approach could be utilized as a persuasive basis for the classification of the starch fractions. The structure of amylosucrase-modified starches was characterized before and after removal of RDS and/or SDS. Branch chains with certain lengths contributed to the organization of each fraction and determined the primary crystallite configuration of amylosucrase-modified starches causing the diversity of digestion properties.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Enzymatic hydrolysis curves of starches were fitted to a logarithm of slope plot. </LI> <LI> The change in rate constant during digestion was observed. </LI> <LI> The individual existence of RDS and SDS was proven in amylosucrase-modified starch. </LI> <LI> The structure of starches was investigated before and after digestion. </LI> <LI> Amylopectin chain length determined the arrangement of each starch fraction. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Preparation and characterization of the inclusion complexes between amylosucrase-treated waxy starch and palmitic acid

김혜인,김하람,최승준,박천석,문태화 한국식품과학회 2017 Food Science and Biotechnology Vol.26 No.2

Amylosucrase-treated waxy corn starch (AS) was produced to extend the chain length of amylopectin to a great extent in comparison to its native chain length. An amylopectin–palmitic acid (PA) complex was prepared by heat-treating (121oC) a starch/PA mixture and its subsequent further incubation (95oC, 24 h); moreover, its structure and digestibility were studied. Unmodified waxy starch could not complex at all, whereas elongation due to amylosucrase modification allowed amylopectin to form a complex with PA to a small extent. Complexation between AS and PA caused a decrease in relative crystallinity. The AS–PA complex displayed an endothermic peak representing type I inclusion complexes rather than type II complexes. The formation of complexes did not significantly affect the in vitro digestibility maintaining the low digestibility of AS resulting from extremely small amounts of complexes and the type of complex.

The structural characteristics of amylosucrase-treated waxy corn starch and relationship between its in vitro digestibility

박천석,박인명 한국식품과학회 2017 Food Science and Biotechnology Vol.26 No.2

The glucotransferase amylosucrase (AS) influences the structural properties of starch, but its precise effects are unclear. The structural characteristics and in vitro digestibility of waxy corn starch modified by AS from Neisseria polysaccharea were examined. AS-treated starch exhibited a higher slowly digestible starch (SDS) fraction, the weak B-type polymorph, lower relative crystallinity, and lower double helix content than those of native starches based on X-ray diffractometry, solid-state 13C CP/MAS NMR, and FT-IR. AS-treated starches exhibited increased proportions of degree of polymerization (DP) 25–36 and DP≥37 chains. Higher SDS and resistant (RS) fractions, higher proportions of DP 25–36 and DP≥37 chains, more double helices, higher relative crystallinity, and less difference between double helix and relative crystallinity were observed for starch treated with 460 U than with 230 U of AS. AS re-built the double-helical and rearranged crystalline structure of gelatinized starch and consequently influenced the SDS and RS fractions.

Molecular Cloning and Functional Expression of a New Amylosucrase from <i>Alteromonas macleodii</i>

HA, Suk-Jin,SEO, Dong-Ho,JUNG, Jong-Hyun,CHA, Jaeho,KIM, Tae-Jip,KIM, Young-Wan,PARK, Cheon-Seok Japan Society for Bioscience, Biotechnology, and A 2009 Bioscience, Biotechnology, and Biochemistry Vol.73 No.7

<P>The presence of amylosucrase in 12 <I>Alteromonas</I> and <I>Pseudoalteromonas</I> strains was examined. Two <I>Alteromonas</I> species (<I>Alteromonas addita</I> KCTC 12195 and <I>Alteromonas macleodii</I> KCTC 2957) possessed genes that had high sequence homology to known amylosucrases. Genomic clones containing the ASase analogs were obtained from <I>A. addita</I> and <I>A. macleodii</I>, and the deduced amino acid sequences of the corresponding genes (<I>aaas</I> and <I>amas</I>, respectively) revealed that they were highly similar to the ASases of <I>Neisseria polysaccharea</I>, <I>Deinococcus radiodurans</I>, and <I>Deinococcus geothermalis</I>. Functional expression of <I>amas</I> in <I>Escherichia coli</I> was successful, and typical ASase activity was detected in purified recombinant AMAS, whereas the purified recombinant AAAS was nonfunctional. Although maximum total activity of AMAS was observed at 45 °C, the ratio of transglycosylation to total activity increased as the temperature decreased from 55 to 25 °C. These results imply that transglycosylation occurs preferentially at lower temperatures while hydrolysis is predominant at higher temperatures.</P>

Effect of short-chain fatty acids on the formation of amylose microparticles by amylosucrase

Lim, Min-Cheol,Park, Kyu-Hwan,Choi, Jong-Hyun,Lee, Da-Hee,Letona, Carlos Andres Morales,Baik, Moo-Yeol,Park, Cheon-Seok,Kim, Young-Rok Elsevier 2016 Carbohydrate polymers Vol.151 No.-

<P><B>Abstract</B></P> <P>Amylose microparticles can be produced by self-assembly of amylose molecules through an amylosucrase-mediated synthesis. Here we investigated the role of short-chain fatty acids in the formation of amylose microparticles and the fate of these fatty acids at the end of the reaction. The rate of self-assembly and production yields of amylose microparticles were significantly enhanced in the presence of fatty acids. The effect was dependent on the length of the fatty acid carbon tail; butanoic acid (C4) was the most effective, followed by hexanoic acid (C6) and octanoic acid (C8). The amylose microparticles were investigated by carrying out SEM, XRD, Raman, NMR, FT-IR and DSC analysis. The size, morphology and crystal structure of the resulting amylose microparticles were comparable with those of amylose microparticles produced without fatty acids. The results indicated the carboxyl group of the fatty acid to be responsible for promoting the self-assembly of amylose chains to form microparticles. The fatty acids were eventually removed from the microstructure through the tight association of amylose double helices to form the amylose microparticles.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Effect of fatty acids on the synthesis of amylose microparticles was investigated. </LI> <LI> Short-chain fatty acids significantly increased production yield of microparticles. </LI> <LI> The carboxyl group of the fatty acid was found to be responsible for this effect. </LI> <LI> Fatty acids were ejected from the microparticles at the end of the self-assembly. </LI> <LI> Physicochemical properties of the microparticles were unaffected by the fatty acids. </LI> </UL> </P>

Enzymatic Synthesis of Polyphenol Glycosides by Amylosucrase

박현수(Hyunsu Park),최경화(Kyoung-Hwa Choi),박영돈(Young Don Park),박천석(Cheon-Seok Park),차재호(Jaeho Cha) 한국생명과학회 2011 생명과학회지 Vol.21 No.11

재조합 아밀로수크라아제의 폴리페놀 배당체를 합성하는 능력을 검사하였다. 이 효소의 효소작용 특성에 근거하여 설탕을 기질로 사용하였으며 21 종류의 각기 다른 폴리페놀 화합물들이 수용체로 사용되었다. 당 전이 반응은 사용한 폴리페놀에 따라 하나 또는 두 개의 주요 폴리페놀 배당체를 합성하였다. 합성된 폴리페놀 배당체들은 박막 크로마토그래피법을 이용하여 확인되었고, 새로이 합성된 배당체의 구조는 당 전이 작용 특성에 근거하여 예측되었다. 수용체로 가능한 폴리페놀의 구조적 특징들이 평가되었으며, 이러한 결과는 Deinococcus geothermalis 유래 아밀로수크라아제가 식품, 화장품, 및 제약산업에서 높은 잠재성을 갖는 폴리페놀 배당체의 효소적 합성에 매우 효율적인 촉매로 활용될 수 있다는 것을 보여준다. The capability of synthesizing polyphenol glycosides was examined using recombinant amylosucrase from the hyperthermophilic bacterium Deinococcus geothermalis. Based on the action mode of amylosucrase, sucrose and twenty-one polyphenols were used as a donor and acceptors respectively. The transglycosylation reaction by amylosucrase produced one or two major polyphenol glycosides depending on the type of polyphenols used. The synthesized polyphenol glycosides were detected by thin-layer chromatography. The structures of the newly synthesized polyphenol glycosides were predicted based on the transglycosylation mechanism of the enzyme. According to the acceptability of the polyphenols, the structural characteristics of polyphenol as an efficient acceptor were evaluated. The results indicate that amylosucrase is an efficient catalyst for the enzymatic synthesis of polyphenol glycosides, which have high potentials in food, cosmetics, and pharmaceutical industries.

Bioconversion of Piceid to Piceid Glucoside Using Amylosucrase from Alteromonas macleodii Deep Ecotype

( Hyun Su Park ),( Ji Eun Kim ),( Ji Hae Park ),( Nam In Baek ),( Cheon Seok Park ),( Hee Seob Lee ),( Jae Ho Cha ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.12

Resveratrol, or its glycoside form piceid, is a dietary antioxidant polyphenolic compound, found in grapes and red wine that has been shown to have protective effects against cardiovascular disease. However, very low water solubility of the compound may limit its application in the food and pharmaceutical industries. The amylosucrase (AMAS) of Alteromonas macleodii Deep ecotype was expressed in Escherichia coli and showed high glycosyltransferase activity to produce the glucosyl piceid when piceid was used as an acceptor. The conversion yield of piceid glucoside was 35.2%. Biotransformation using culture of the E. coli harboring the amas gene increased the yield up to 70.8%. The transfer product was purified by reverse phase chromatography and recycling preparative HPLC, and the molecular structure of the piceid glucoside was determined using NMR spectroscopy. The piceid glucoside was identified as glucosyl-α-(1→4)-piceid. The solubility of glucosyl piceid was 5.26 and 1.14 times higher than those of resveratrol and piceid, respectively. It is anticipated that dietary intake of this compound is more effective by enhancing the bioavailability of resveratrol in the human body because of its hydrophilic properties in the intestinal fluid.

Functional Expression of Amylosucrase, a Glucan-Synthesizing Enzyme, from Arthrobacter chlorophenolicus A6

( Seo Dong Ho ),( Jong Hyun Jung ),( Hyun Chang Choi ),( Hyun Kuk Cho ),( Hee Hang Kim ),( Suk Jin Ha ),( Sang Ho Yoo ),( Jae Ho Cha ),( Cheon Seok Park ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.9

A gene (acas) designated as α-amylase was cloned from Arthrobacter chlorophenolicus A6. The multiple amino acid sequence analysis and functional expression of acas revealed that this gene really encoded an amylosucrase (ASase) instead of α-amylase. In fact, the recombinant enzyme exhibited typical ASase activity by showing both sucrose hydrolysis and glucosyltransferase activities. The purified enzyme has a molecular mass of 72 kDa and exhibits optimal hydrolysis activity at 45oC and a pH of 8.0. The analysis of the oligomeric state of ACAS with gel permeation chromatography revealed that the ACAS existed as a monomer.

Enzymatic Synthesis of Resveratrol α-Glucoside by Amylosucrase of Deinococcus geothermalis

( Keumok Moon ),( Seola Lee ),( Hyunsu Park ),( Jaeho Cha ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.12

Glycosylation of resveratrol was carried out by using the amylosucrase of Deinococcus geothermalis, and the glycosylated products were tested for their solubility, chemical stability, and biological activities. We synthesized and identified these two major glycosylated products as resveratrol-4′-O-α-glucoside and resveratrol-3-O-α-glucoside by nuclear magnetic resonance analysis with a ratio of 5:1. The water solubilities of the two resveratrol-α-glucoside isomers (α-piceid isomers) were approximately 3.6 and 13.5 times higher than that of β-piceid and resveratrol, respectively, and they were also highly stable in buffered solutions. The antioxidant activity of the α-piceid isomers, examined by radical scavenging capability, showed it to be initially lower than that of resveratrol, but as time passed, the α-piceid isomers’ activity reached a level similar to that of resveratrol. The α-piceid isomers also showed better inhibitory activity against tyrosinase and melanin synthesis in B16F10 melanoma cells than β-piceid. The cellular uptake of the α-piceid isomers, which was assessed by ultra-performance liquid chromatography (UPLC) analysis of the cell-free extracts of B16F10 melanoma cells, demonstrated that the glycosylated form of resveratrol was gradually converted to resveratrol inside the cells. These results indicate that the enzymatic glycosylation of resveratrol could be a useful method for enhancing the bioavailability of resveratrol.