α-Glucosidase Inhibitor (AGI) from α-Glucosidase Inhibitor (AGI) from α-Glucosidase Inhibitor (AGI) from Natto : Production and Applicationα-Glucosidase Inhibitor (AGI) from Natto : Production and Applicationpppp

Sera Won Shinji Yamamoto*and Hae ik RheeSera Won, Shinji Yamamoto*and Hae-ik Rhee

Division of Biotechnology, Kangwon National University, Koread *N Bi E i Okand *New Bio Enterprise, Okayama

Digestion of Carbohydrates

P l Di li MPoly-saccharide

Di, oligo-saccharide

Mono-saccharideα-glucosidaseα-amylase

α-amylaseStarch

glucosemaltose

glucosemaltaseα amylaseStarchmaltotriosedextrin

glucose

sucrosefructose,glucosesucrase

lactosegalactose,

llactase

glucose

Normal absorption of disaccharide

di h id disaccharide +

enzymehydrolysis absorption of

monosaccharide

Inhibition of α-glucosidase by AGI

inhibitor +

enzymeinhibition of

hydrolysis & absorption

Cheong-guk-chang

Characteristics of DC-15 Gram staining +

Shape rod

Nitrate reduced to nitrite +Formation of Indole -Catalase +

Size 0.6 ~ 0.7 ×1.6 ~ 1.9 ㎛

Spore O

Catalase +Hydrolysis of starch +Hydrolysis of casein +p

Spores position endospore

til

yd o ys s o caseUtilization of propionate -Acid production from

Motile +

Growth temperature 25 ~ 40℃Glycerol +L-Arabinose +D Xylose +Growth in NaCl 7% +

Aerobic growth +

D-Xylose +D-Galactose +D-Glucose +Aerobic growth

Anaerobic growth -D-Mannitol +D-Maltose +

Voges-Proskauer test +D-Sucrose +D-Lactose +

Electron microscopy of DC-15

DC-15 Natto

Distribution of α－glucosidase inhibitory activities

Sample UnitX103/ｇp ｇ

Freeze dry 21,000Spray dry 23 000

DC-15 nattoSpray dry 23,000wet, 24hr 7,400wet 36hr 12 800wet, 36hr 12,800wet, 48hr 17,800

1

Commercial

1 none2 none

Commercial natto 3 none

4 none5 none

200

mg/

d)

160co

se (m

120`

d g

luc

80`

Control

Bloo

d 40

0

Control DC-15 natto

Time (min)

00 30 60 90 120

Time (min)

Eff t f DC 15 tt bl d l l lEffect of DC-15 natto on blood glucose levels

AGI activity in DC 15 natto ramenAGI activity in DC-15 natto ramen

S l U it 103/ Natto(g)/Total activity (unit x 103 ）

Sample Unit x 103/g Natto(g)/Flour 100g

Estimated Actual

DC-15 natto(Spray Dry） 20,850 0 - -(Spray Dry）

Ramen 1 2 41 700 37 000Ramen 1 2 41,700 37,000

R 2 3 62 550 60 000Ramen 2 3 62,550 60,000

160d) Control 160

120(mg/

d DC-15 natto

120

80ucos

e

80

40od g

lu

40

0

Bloo

0 0 30 60 90 120 150Time (min)

Effect on blood glucose levels after ingesting ramen containing DC-15 natto

Diet composition p

Ingredient (g/kg) Control DC-15

Casein 204 204Casein 204 204Sucrose 699 699Corn oil 50 50Mineral mix 35 35Vitamin mix 10 10Choline chloride 2 2Commercial natto 15 -DC-15 natto - 15DC 15 natto 15

) 160

ght (

g)120

Control DC-15 natto

y w

eig

80

e bo

dy

80

40

crea

se 40

0

D

Inc 0

0 5 10 15 20 25

Days

Eff t f DC 15 tt b d i ht f tEffect of DC-15 natto on body weight of rats

Placebo DC-15 NattoPlacebo DC 15 Natto t (

kg) 1

0 0

1w

eigh

t

-1

2

-1

2

d bo

dy -2

-3

-2

-3

crea

se -4 -4

-5-5

In 20100 30 0 10 20 3040 40

Days Days

B d i ht i i f 30 dBody weight gain in women for 30 days

Conclusion

We isolated microorganism producing α glucosidase

Conclusion

We isolated microorganism producing α-glucosidaseinhibitor from Cheong-guk-chang which is traditionalfermented soybean food in Koreafermented soybean food in Korea.

It is identified Bacillus subtilis and named DC-15.

After ingesting DC-15 natto, increase of blood glucose isrepressed.

There is significant body weight decrease after givingDC-15 nattoDC 15 natto.

DC-15 natto can be used as a functional food withoutt ti d t tiextraction and concentration.