In vitro Analysis on Inhibitory Activity of Amylolytic Enzymes in Deca# ee Bean Extracts and Contributions of Chlorogenic Acids

Transcription

1 336 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /0, No. 0, --0-., (,**3) 28 UCC R& D In vitro Analysis on Inhibitory Activity of Amylolytic Enzymes in Deca# einated Green Co# ee Bean Extracts and Contributions of Chlorogenic Acids Yuji Kamitani, Kazuya Iwai, Taiji Fukunaga, Ryotaro Kimura and Osamu Nakagiri R& D Center, UCC Ueshima Co# ee Co, Ltd., - +. Zushi, Takatsuki-Shi, Osaka /03 **-0 Extract of deca# einated green co# ee beans (EDGCB) was prepared by supercritical CO, treatment and alcohol extraction and contained -2 (w/w) chlorogenic acids (CQAs, consisting of 2 kinds of chlorogenic acid isomers). The e# ects of CQAs in EDGCB on inhibitory contribution of amylolytic enzymes ( a-glucosidase and a-amylase) were examined in vitro. The activity of these amylolytic enzymes was inhibited depending on the EDGCB concentration, and CQAs significantly contributed by /. 2. Comparison of CQA isomers showed that inhibitory activities of di-ca# eoylquinic acids (-/, -dicqa, -., -dicqa and./, -dicqa) were the strongest, followed by those of ca# eoylquinic acids (--CQA, /-CQA and.-cqa) and feruloylquinic acids (/- FQA and.-fqa) in a-amylase and sucrase. -., -dicqa was the main active component in EDGCB for inhibiting maltase. The number of ca# eoyl groups and the binding position for quinic acid significantly a# ected the inhibitory activity of CQAs. The e# ects of EDGCB on the inhibitory activity of a-gi (acarbose or voglibose) in vitro and on the postprandial blood glucose level were next examined using rats. Addition of EDGCB to the reaction mixture with a low concentration of a-gi showed additive inhibitory e# ects of each enzyme. Moreover, EDGCB prevented the increase in blood glucose level in vivo. These results suggest that EDGCB is useful for prevention of postprandial hyperglycemia. (Received Nov. +-,,**2 ; Accepted Mar. 0,,**3) Keywords : extraction, deca# einated green co# ee beans, chlorogenic acids, amylolytic enzyme, inhibitory activity, postprandial blood glucose : ao ao + -, -. /0 1 2 CQA FQA dicqa 2 ao ao +* ++ +,., /03 ** Corresponding authoryuuji-kamitani@ucc.co.jp 3 + CO 1*./ MPa 0*,

2 29 : ao ABCDAB+** Indonesia AP-+ A:, B: Sephadex C : D : LH-,* GE ao CQAFQA dicqa HPLC p-nitrophenyl-a-d-maltoside G, - PLC/0+ --CQA, /-CQA,.- pnpsigma p-nitrophenol CQA/ -FQA.-FQA -,/-dicqa -,.-dicqa.,/-dicqa.*/ nm +* L,* mm ph 1*. +* - ao SIGMA,* L,* mm HPLC ph 1.* +0* L -/ HPLC -*,* mm G -pnp +* L : -/ 0* : Inertsil ODS mm +**.* : A *., 2*,* B : * min A +** ao // B *./ 0* min A // B./ 0*0/ min A +** B * : -,/ nm : +.* +- mlmin. ao A B A +** / -CQA : +.**, --CQA : +.*1,.-CQA : +.*1, /-FQA : +.*.,.- A : FQA : +*.., -/, -dicqa : *3., -., -dicqa : *3.,./, -dicqa : B : * / SIGMA, g /0mM ph 0.*./ ml / *** rpm IC +** ao IC IC,*. +1 / ao +0 / a-gi L /0 mm : *, gml ph 0.* L L, :,. 10- gml, L : *. ++ *... gml -1 -* *+. */. +*. +/. mgml. 2** L,** L ao +* 0* +2* gml C. 0 Toronto Research Chemicals Inc. ao 0 SPF, SD 1 1 0,** mgml,

3 338 /0 0,** Table + Chlorogenic acids content of EDGCB CQA, FQA dicqa Content ( w/w) --CQA /4+,.-. + IC : +.*+ mgml -/. + IC : *.,0* mgml /-CQA +.42 ao.*. + IC : *. +2, mgml./. + IC :.-CQA 14*- *.*-. mgml /-FQA -42. CQA, FQA dicqa.-fqa +4, IC : *. 2*2 mgml /0. 1 IC : *.*.. -,/-dicqa +4/3 -,.-dicqa,4+* mgml dicqa CQA, FQA..,/-diCQA CQA, FQA -4*. -,4+ Table, 2 dicqa 041- ao /-CQA EDGCB : Extract of deca# einated green co# ee beans +3. 3,-.- /-CQA mg ml,. +* +0 -,.-dicqa * mlkg Table, -., -dicqa : +01 mgkg, * mgkg, + * mgkg, : * mgkg 3 -* 0* +,* dicqa : /,, -tricqa +, ao Tukey-Kramer / + dicqa -/, -dicqa, -., -dicqa,./, -dicqacqa --CQA, /-CQA,.-CQAFQA / -FQA,.-FQA + CQA, FQA -,. + dicqa CQA /-CQA FQA dicqa CQA /+,.,.-CQA 1*-., /-FQA 2/ ,.-FQA +,3., -/, -dicqa +/3., -., -dicqa,+*../, -dicqa -*.. Table + : **-. : +** g -+. g *,. +*.. g +*., g.*,,. g *,. g ***-. g :,*0++*---, a-gi ab ao c a-gi Fig. + b

4 31 : 339 Fig. + Inhibitory e# ects of EDGCB and a-gi on Maltase, Sucrase and a-amylase activity Table, The comparison of contribution ratio and inhibitory activitiy of amylolytic enzymes Maltase Sucrase a-amylase IC (mg/ml) Contribution( ) IC (mg/ml) Contribution( ) IC (mg/ml) Contribution( ) EDGCB CQA, FQA fraction dicqa fraction * *4,,2 *42* *+.-4+ *4+2,.*4. *4*.. /041 *4,0* -/4+ *4*-../4+ --CQA /-CQA.-CQA /-FQA.-FQA -,/-dicqa -,.-dicqa.,/-dicqa *42+0 *4133 *42+., *4,/, *4*,* *4+/2,4-/ ,- +4*- *4--,4, * *+ *432* * *4-., *4,.- *4,30 14* ,41/ *43, *4+02 *4+./ *4+0, *4.20 *4../ *4* *4*,1 *4*-2 043/, * *400 14, CQAs ,4-2/42 IC : Concentration which inhibits amylolytic enzyme activity to

5 340 /0 0,** a-gi in vitro a-gi a-gi Table - a-gi +. a-gi mgkg * mgkg + * mgkg -* Fig., * mgkg * mgkg * mgkg -* 0* Fig. - in vitro in vitro ao ao in vivo,* CQA FQA dicqa in vitro in vivo in vivo Na. in vitro Table - Inhibition of Maltase, Sucrase, and a-amylase activity by a-gi and EDGCB mixture Maltase activity a-gi and EDGCB Inhibition ( ) Acarbose *4,- g/ml.24, Acarbose *4,- gedgcb *4+ mg/ml /04* Acarbose *4,- gedgcb *4/ mg/ml 0141 Acarbose *4,- gedgcb +4* mg/ml 1.42 Acarbose *4,- gedgcb +4/ mg/ml 134, Acarbose 04. g/ml 2+4, Acarbose 04. gedgcb *4+ mg/ml 2-4* Acarbose 04. gedgcb *4/ mg/ml 2/41 Acarbose 04. gedgcb +4* mg/ml 214, Acarbose 04. gedgcb +4/ mg/ml 2143 Voglibose *4++ g/ml.343 Voglibose *4++ gedgcb *4+ mg/ml 0-41 Voglibose *4++ gedgcb *4/ mg/ml 124. Voglibose *4++ gedgcb +4* mg/ml 2/4+ Voglibose *4++ gedgcb +4/ mg/ml 234- Voglibose *4.. g/ml 2+4. Voglibose *4.. gedgcb *4+ mg/ml 204, Voglibose *4.. gedgcb *4/ mg/ml 3*4+ Voglibose *4.. gedgcb +4* mg/ml 3+41 Voglibose *4.. gedgcb +4/ mg/ml 3-4, Sucrase activity a-gi and EDGCB Inhibition ( ) Acarbose,41 g/ml /+4* Acarbose,41 gedgcb *4+ mg/ml /34/ Acarbose,41 gedgcb *4/ mg/ml 1*4+ Acarbose,41 gedgcb +4* mg/ml 1/4* Acarbose,41 gedgcb +4/ mg/ml 1340 Acarbose 0- g/ml 2,4- Acarbose 0- gedgcb *4+ mg/ml 2.42 Acarbose 0- gedgcb *4/ mg/ml 204/ Acarbose 0- gedgcb +4* mg/ml 2141 Acarbose 0- gedgcb +4/ mg/ml 2240 Voglibose *4++ g/ml /-43 Voglibose *4++ gedgcb *4+ mg/ml 0-4* Voglibose *4++ gedgcb *4/ mg/ml 1,4+ Voglibose *4++ gedgcb +4* mg/ml 104* Voglibose *4++ gedgcb +4/ mg/ml 2+4. Voglibose *4.. g/ml 2+43 Voglibose *4.. gedgcb *4+ mg/ml 2/4- Voglibose *4.. gedgcb *4/ mg/ml 2042 Voglibose *4.. gedgcb +4* mg/ml 224* Voglibose *4.. gedgcb +4/ mg/ml 2243 a-amylase activity a-gi and EDGCB Inhibition ( ) Acarbose 0* g/ml /-4- Acarbose 0* gedgcb *4+ mg/ml 0141 Acarbose 0* gedgcb *4/ mg/ml 1-4- Acarbose 0* gedgcb +4* mg/ml 2040 Acarbose 0* gedgcb +4/ mg/ml 3+4. Acarbose +2* g/ml 2343 Acarbose +2* gedgcb *4+ mg/ml 3-42 Acarbose +2* gedgcb *4/ mg/ml 3141 Acarbose +2* gedgcb +4* mg/ml 3240 Acarbose +2* gedgcb +4/ mg/ml 3241

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