TAP CHl CONG THirONG
TOI l/U DIEU KIEN BIEN TINH TINH BOT GAO TAO TINH BOT KHONG TIEU HOA BANG ENZYME
• vo MINH HOANG - N G U Y I N DLfC TOAN
T 6 M TAT:
Bai vie't nghien effu cdeh td'i ffu hda eac thdng so'cua qud trinh thuy phan nhdm tao hdm Iffdng tinh bot RS cao nha't bdng enzyme maltogenie amylase. Ke't qud phan tich cho tha'y, phifdng trinh hoi quy da thffc bae hai thu dffde cd y nghTa {p <0,05). Lackof fit/j-value >0,05 chi ra rdng. md hlnh phii hdp cho ta't ca dff lieu. Dp tin cay cua md hmh R- = 0,998, do tffdng thieh cua mo hinh Q- = 0.812 cho tha'y md hlnh tiTdng thich vdi ket qua thffc nghiem va c6 do tin cay cao. Dieu kien to'i ffu eho qud irinh thuy phand nhiet do 60,015"C, thdi gian 6,679h va nong do enzyme 29.352U/ml cho ham Iffdng tinh bdt khdng tieu hda eao nha't ia 6,04%.
Tiif khoa: Bien tinh tinh bdt, tdi ffu hda, tinh bot khdng thuy phan (RS).
1. Dat %a'n de
Tinh trang thffa can va beo phi dang tang len vdi td'c do bao ddng khdng nhffng d cac quo'c gia phat trien ma 8 ca cde qudc gia dang phdt tnen.
Nguyen nhan cd ban cua thffa can. beo phi la sff ma't can bang ndng Iffdng giffa Iffdng ealo tieu thu
\a kfdng ealo tieu hao. Md khau phan an cQa chiing ta ham Iffdng tinh bdt tieu hda nhanh chieni t\ le qua eao. dieu nay dan de'n mot trong nhffng nguyen nhan ga> ra benh nay.
De giai quye't nhffng van de na>. nhieu nha khoa hoc dd nghien cffu tinh bdt cd linh chat hda h' Id'l hdn. Theo Englysl \h cdc cdng sff (1992). dd'i
\di mue dich dinh dffdng. tinh bdt irong ihifc pham dffdc phan loai thanh tinh hot tieu hda nhanh IRDS - Rapidly Digestible Starch), tinh bdt tieu hda chdm (SDS - Slowly Digestible Starch) va tinh bdt khdng tieu hda (RS - Resi.stant Starch) dffa theo id'cdd giai phdng glucose \a ha'p thu cija nd trong
dffdng tieu hda Va RS la tinh bdt khdng bi th phan sau 120 phiit xff ly vdi he enzyme amyl;
tuye'n tuy nhu'ng bi len men it hoae nhieu bdi he smh vat dffdng rudt san sinh ra cdc acid beo ma ngdn cd Idi eho di/dng rudt va gdp phan bt) su them ngudn ndng lu'dng cho cd the. Do dd, ddng vai trd nhu"cha't xd nit ngan thdi gian tieu li cua thffc pham va giam sd phat trien cua cae I thu'dng ve du'dng mdt [2 - 6]. Ve gia tri nang Iffc Clia RS trong thffc pham da dffdc tinh todn khoa 2.2 kcal/g [7). Dffa tren ndng Iffdng RS da dffde xua't nhff mdl thanh phan trong ehe dp an cd l gilip giam can [8]. phdng ,su' suy giam ldp nhdy t v^ rupl ke't, gidm nguy cd ma'c ung thi/rupt ket hd trd kiem soat benh ddi thao dudngld. 9 - i giam cholesterol trong mdu. giam chi so' dffo huye't, tang hap thu chat khoang [5. 9. I2|.
Mue dich sff dung enzyme malto<!enic amyl;
Ihuy phan tinh bdt nhdm tao ra loai tinh bdt e
364 So 9-Thang 5/2020
HOA HOC-CONB NGHE TKKC PHAM
bien tinh cd dp phan nhanh cao. ca'u tnic tinh the iang de lam cham kha nang tieu hda. Hieu qud thuy phan tinh bdt ciia enzyme phu thupc vao nhieu Jicu kien. dae biet la nhiet dp. thdi gian va nong dp enzyme thijy phan. Vi vay, nghien effu nay dffdc thffc hien nhdm muc lieu td'i ffu hoa eae thdng so Clia qua trinh ihijy phan nham tao ham lifting tinh hot RS cao nha't.
2. Phffdng phap nghien cffu 2.1. Nguyen U^u vd hda chat
Bdt gao dude lam tff lua gao (Oryza saliva L.) dffdc mua tff ed sd Lpc Sdnh. a'p Tan My, xa Tan Phii Trung. huyen Chdu Thanh - tinh Ddng Thdp.
Dd a'm 14.59%: protein ().657f; lipit 0.3%: gluxil 81.37';f; Id'ng sd vi sinh vat hie'u khi 5xl0Uvsv/g).
Mjltogenie amylase (TQ), a-amylase (hang Novozymes), nguon gd'c tff vi khuan Bacillus .\iibiilis: solube .starch, maltose, D-glueose. 3.5 dinitrosalicylic acid (DNS).
2.2. Phifcfng phdp nghien ci'fu
Trong nghien effu nay tde gia thffc hien 2 thi nghiem chinh: (1) thie't kemd hlnh hda, td'i im hda:
l2i Xiic djnh ham Iffdng RS tai dieu kien td'i ffu.
2 2 1. Thie'l ke lhi nghiem nid hinh hda vd ic'ii lai hda bang phifcfng phdp he null ddp dng
Md hlnh hda difde thie't ke bdi phan mem Design-Expert vdi 3 yeu td': nhiet dd. thdi gian va nong dp enzyme vdi ham muc lieu Id dffdng kliff Jan ra dat thd'p nha't theo mong ddi. (Bang 1)
Bang 1. Cac mCfc dp dUdc md tioa cua mdi bien trong md liinh Box - Behnken
MffcdO
um do,
°c • XlThdi gian, h X2 Nong do enzyme, U/ml I X3
Trong nghien cffu nay. bd tri thi nghiem md hlnh hoa dffdc thffc hien dtJa theo md hlnh Bo\-Behnken
\di3 thdng sd nhiet dp (XI), thdi gian (X.) va nong do enzyme (X;) la cde bie'n ddc lap va ham Iffdng dffdng khff (Yniin) la ddp ffng phu thude. phffdng trinh hoi quy da thffc dffdc dp dung de xual nhff phffdng tnnh:
= /?o + ^ A ^ . + ^ A . J f , ' + 2 ^ ^,,X,X,^
Trong dd: "i" Id bie'n phu ihudc. (^Q la he sd chdn: /?,• la he sd phffdng irinh bae 1. /J„ la he so phffdng trinh bae 2 cua bie'n \i. fij. la he so' tffdng tac va e la sai sd'ngau nhien.
2.2.2. Xdc dinb ham lUi/ng RS lif mdu ldi ini Tff md hlnh todn hpc iinh todn dffdc thdng sd tdi ffu cho dieu kien bie'n tinh Mau tinh bdt td'i ifii sau dd se dffde ddnh gid ham lu'dng RS vd so sdnh vdi mau thffc te trong ciing dieu kien. Ham Iffdng RS trong cac mau nghien cffu nay du'de xac dinh dffa theo phffdng phdp Englysi et al. 1992 vdi mdl so'dieu chinh. Cdng thffc \dc dinh ham Iifdng RS nhff sau:
TS =(TG-FG)^0.9 RSD = (G20-FG)*0.9 SDS = (GI20-G20)*0.9 RS =TS-(RSD + SDS)
Trong dd: FG la ham kfdng glucose thdi gian bat dau ihiiy phdn (0 phut): G20 la ham lu'dng glucose sau thuy phan 20 phut: G120 la ham Iffdng glucose sau thuy phan 120 phiit: TG la ham lu'dng glucose tdng.
2.2 3. Phdn tich ihe'nig kc
Ke't qua difdc \ff !>' bang phan mem thd'ng ke SPSS Statistics 20 I IBM) Ke't qua thi nghiem ldi ffu hda difdc xff ly bang Design-Expert 11.0 4 0 (Stai-Ease)
3. Kd't qua thao lu§n 3.L Me') hinh hda thUc nghiem
Md ta qua trinh xff ly bang md hlnh thifc nghiem bae 2 Box-Behnken. Ma Iran thffc nghiem va ke't qua thffc nghiem nhd (Bang 2).
Ket qua tai Bang 3 cho tha'y md hinh ed y nghia vdi p-\alue < 0.0001. Lack of fit p-value > 0,05 dieu nay chi ra rdng md hlnh phii hdp eho tat ca dff lieu. Dp tin eay ciia md hinh R- = 0.998, dd tffdng thich cua md hinh Q- = 0,812 eho tha'y md hlnh tffdng thich \6'\ ke't qua thffc nghiem va cd dp tin ea\ cao. R- Adj. = 0.995 tu'dng dffdng vdi R- chu tha'\ cdc \e'u idkh.io sdt da giai thich phan ldn ke't qua thi nghiem. R- dff doan la thffdc do mffc dp phii hdp cua md hlnh dff dodn. trong pham vi 0.20 ciia R- -Adj la phu hdp.
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TAP CHi CdNG THIfONG
Bang 2. Gid tri quan sdt vd gia hi dff dodn trong mo tiinti tido
Mau
N1 N2 N3 N4 N5 N6 N7 N8 N9 N 1 0 N i l N12 N13 N14 N15
TTiiltLr c h a y m a u
6 9 14 11 4 5 7 13 3 8 15 1 10 12 2
XI
1 0 0 0 1 1 1 0 1 1 0 1 0 0 1
X2
0 1 0 1 1
°
0°
1 0 0 1 1 1 1X 3
• 1
-1 0 1 0
• 1
1 0 0 1 0 0 .1 1 0
D u d n g khO (ijmol/ml)
Q u a n s a t 1,49 1.35 1,10 1,29 1.42 1,37 1.36 1.09 1,34 1,29 1.08 1.46 1.34 1.15 1.45
D u d o a n
1.60 1,35 1.09 1.30 1.42 1,37 1,35 1.09 1.34 1.29 1.09 1.46 1.34 1,16 1,45
Bang 3. Ptidn tich thong ke onovo ctio dffdng ktiuf Nguon
M o d e l 1 X ,
X, X j 1 ' ^ l ^2
X,X3 X;X3 X;
x;
X- Residual Lack of Fit
SSa 0 2671 0 0018 0,0113 0.0265 0.0020 0.0090 0.0042 0,1648 0,0499 0.0215 0 0004 0 0002
d1»
9 1
5 3
MSc 0 0 2 9 7 0 0 0 1 8 00113 0 0265 0.0020 0.0090 0 0042 01648 0-0499 0.0215 0.0001 0.0001
F-value 329 78 20.00 125.00 293.89 22.50 100.28 46 94 1830 83
554.42 238.53
0 8333 [ l v a l u e
< 0 0 0 0 1 0 . 0 0 6 6
< 0 0 0 0 1
< 0.0001 0-0051 0.0002 0.0010
< 0 0001
< 0 0 0 0 1
< 0 0 0 0 1
0 . 5 8 5 9
Y ngtiTa
K h o n g y n g t i l a
3 6 6 So 9 - Thang 5/2020
IdA IOC • CONE NOHE IHIfC PIAM
Nguon Pure Error R- Adjusted R' Predicted R=
Adequate Precision
SSa 0.0002 0.9983 0.9953 0.9834
df 2
MSc 0 0001
F-value
- - -
p-value
52.6080 .
Ghi chli:" long bhib phUcfng,'' bgc tu do. ' trung hinh binh phifcfng Phffdng trinh hoi quy cd dang:
Y = 1.09 + 0.0151 X| - 0.0375X^ - 0.0.';75X, + 0.0225X|X2 - 0.0475X|Xi - 0.032.^X2X3 + 0.2112X1 + 0,1162X5 + 0.0762X^, Tff phifdng trinh todn hpc. chiing tdi td'i ffu hda qud trtnh thuy phan ma tai dd Iffdng du'dng khff tao ihanh la tha'p nha'l iheo mong mud'n. (Bang 4).
3.2. kiem tra cdc dieu ki^n td'i tfu Mau sff dung cho thi nghiem kicm tra tai dieu kicn Id'l \iu: 100ml dung dieh ho tinh bdt 5%(w/w) lai nhieldp60.0l.')"C. thdi gian 6.679hva nong do en/) me 29.352U/ml. Kel qua thu dffdc nhif Bang 5
\di 3 lan lap lai.
Theo Englysl ct al. (1992) Uong 20 phiil dau
(G20) dnh bdt RDS dUde ihuy phdn irffde. Qua Bang 5 cho iha'> lffdng glucose dUde giai phdng lai thdi diem G20 d mau tdi ffu thap hdn .so mau tff nhien. Dieu naj cho thd'y ed sff bie'n ehuyen thanh phan RDS trong tinh bdt tff nhien sang cdc dang khde khd thuy phan hdn, Sau 120 phut tinh bdi RDS \ a tinh bdt SDS ,se dffdc thily phan hodn loan thanh glucose. Tai thdi diem G120 sff chenh leeh hdm Iffdng difdng khff giai phdng giffa 2 mau chi
\ao khoang 0.65pniol/ml tha'p hdn 8 lan so vdi sif chenh leeh tai thdi diem G20. Nhu" vay, linh bdi iff nhien sau khi \ff ly bdng enzyme maltogenie amylase thi thanh phan RDS chuyen hda thanh SDS\aRS.
Bang 4. Ke't qua tdi L/U hoa
, Ham IUdng dffdng khCi (Imol/ml) Nhi^tdOCC) Thdi gian (h) Mautoiuu 60,015 6.679 29,352
Nong dO enzyme (U/ml) 1,074
Hinh 1: Od thj be mat dap (ing cua ham IUdng difdng khCi
(a) Tcic dgng ciia nhiel do vdt llidi gicni (leu gid tri ndng do eiKvnie Id 29.3.~'2r/mli (bi Tac dgng ciia nhiet do vdi nchig do enzyme (igi llidi gian 6.679hl
(Cl Tde ddng ciia thdi gian vdi nchig de} enzynw (igi mdc nhiel eld 60.015 C)
So 9-Thang 5/2020 367
TAP CNJ CdNG THtrfTNE
Kct q u a tai B a n g 6 c h o thay gid iri trung binh ciia h a m lu'dng RS d thi nghiem k i e m chffng la 5.S4'>f, Trong khi dd. ke't q u a td'i un h d a cho h a m Iffdng R S dat 6,04% d CLing dieu kien. Q u a ket qua tren cho tha'y c a c thanh phan R D S trong m a u tmh bdt tff nhien sau khi dffdc xff ly bdng maltogenie a m y l a s e thi c h u y e n hda thdnh S D S v a R S .
4. Ke't luSn
Tdi ffu h d a dieu kien thiiy phan tinh bpi cho h a m Iffdng tinh bdt RS e a o nha'l trong qud trinh thuy phan linh hot b a n g e n z y m e maltogenie a m y l a s e dffde \ a \ dffng dffa tren bd' tri thi n g h i e m theo phffdng phdp md hlnh phffc hdp trung t a m cd dp ehinh xac cao, Ke't qua td'i ffu dffdc xff ly d dieu kien nhiet dd 60,015"C. thdi gian thuy phan 6,679h va nong d p e n z y m e 29.3.*i2U/nil cho h a m Ufdng RS c a o nha'l la 6.04'^f •
Bang 5. Hdm lffdng glucose d cdc khoang ttidl gion ktidc ntiou
Mali
T61 uu Tu ntiien
FG 0.13 ±0.006 0.06 ± 0.004
Glucose giai phong (pmol/ml) G20 , G120 18.17 ±0.258 25,66 ±0.458 23,53 ±0.341 26.31 ±0.161
TG 27,31 ± 0.5:
26.74 ±0,1f
Bong 6. Bong ket quo ttii ngtilem kiem ctiifng
1 !
' Niiietdo
! xilly, "C Bien Thoi gian, h
Nong do 1 enzyme, U/ml Ham lupng RS,
%(w/w)
Mo hint! Mo hinti toi uu ttiMc te
60.015 60
6.679 ' 6.5 29,352
6,04" ±0,514 30
6,84" ±0.151 Mail tu nhien
1.62'±0,28
Ghi clui: a. b (p < 0,05}: khac biel co y ngliTa, so lieu Id Irung hin ctia 3 ldn lap.
TAI LIEU T H A M K H A O :
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2, D, L Topping and P. M Clifion (2001). Short-chain fatly acids and human colonic function: Roles of resislan slarchandnonsiarchpolysacchandes Physiol Rev..81(3).pp.l03t-l064 DOI: 10,1152/physrev.2001.fil 3 1031.
3 D, Topping. M Fukushima. & .A. Bird. (2003). Rc-isiani siarch as a prebiolic and synbioiic: Siaie of the art Praccechiigs of the Nuiiilicm Society. 62( I), pp. 17 M 76, DOI-10 IO79/PNS2002224
4 M Shiniada. K Mocliizuki. and T Coda (2d<JSi Dietary Re.sistani Starch Reduces Levels of Glucose Depenilcni Insulinoiropic Polypepude mRNA along lhe Jejunum-Ileum in Both Norma! and Type 2 Diabetic Rats Biosci. eifWf/i/io/.Bfw/jt'Hi.. 72(8). pp22()6-2209. DOI: I0.1271/bbb.80185.
5. E Fuenies-Zaragoza. E. Sanchez-Zapaia. E. Sendra. E Sayas. C. Navarro. J. Femdndez-Ldpez. J. A. P^rez Alvarez (2011). ReMsianl starch as prebiolic: A review. Stai-cli/Staerkc. 63(7). pp. 406-415 DOI lO.IO02/slar.20IO00099.
6- E. Sanchez-Zapata. -M. \ luda-.Manns. J, Fernandez-LOpez. and J, A, Perez-Alvarez (2015). Resisianl starch a' lunciional ingredicni Poly.saccharide\ Bioaciiviiy Bioiechnol.. pp.1911-1931. DOI: n).l(X)7/y7K-3-3]9-j(i298 t)_34.
7, K. M B c h a l l a n d J C Howe 11996). Resistant starch as energy. X/^/H. OJ//. Ai/fr. 15(3). pp 248-254. DOI 10.1080/07315724 1996.10718595
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IDA I O C ' CONG NGHE THOC PHAM
S, L, C. TapinV (2004). Diet and metabolic syndrome Wliere does resistant starch fit in?. J. AOAC Int. S7( 1) nn 756-760, DOI: (nt'u co)
9, .\. P. Nugent (2005). Health properties of resistant slarch. Niiir. Bull.. M I), pp. :7-';4 DOI- 10 111 l/j 1467- .1010.2005 00481.x.
10. J Nissar, T. Ahad, H, R, Naik, and S. Z. Hussain (2017). "Resislanl Slarch- Chemisu^' and Nutritional propettii:^. ItnernotionalJanrnal oJ Food Science and .Mitrttion. 2l6].pp.95-]0S.
11. E. Jyoshna and T. V Hymavalhi (2017). Review of studies on effect of resistant starch supplementation on
^\\iat^litniinni\in.JotintalofPltartnatogniisyandPli\toclietnislf\.6t^).pp.?5:'$
12, M G, Sajilala, R. S. Singhal, and P R. Kulkami (2006). Resistant starch - .A reiiew. Coiniir. Rer. Food Set foodSo/. 5(1), pp. I-17. DOI: tO 111 l/j.l54l-43.')7 20()fi.tb00076 x.
Nuay nhgn bai: 9/4/2020
Ngay phan biC'n danh gia va sufa chifa: 19/4/2020 Ngay cha'p nhan dang bai: 29/4/2020 TItdttii tin ilic liici:
V() MINH HOANG-NGUYEN D C C T O A N
Trung tSni Cong ngh0 Sau thu boach, Khoa Nong nghiep - Thiiy .san Trififng D^i hgfc Tra Vinh
OPTIMIZING MODIFICATION OF RICE STARCH WITH ENZYME TO CREATE RESISTANT STARCH
• VO MINH HOANG
• NGUYEN DUC TOAN Center of Post-harvest Tectinoiogy, Deportinent of Agriculture - Fisheries
Tra Vinh University ABSTRACT:
This study is to find out how to optimize tfie condition of hydrolyzing rice starch with the enzyme maltogenie amylase to produce resistant starch. This study shows that the quadratic polynomial regression equation obtained is significant (p <0.05). Lack of fit p-value> 0.05 indicates that the proposed model is suitable for all data. The reliability of the model R- = 0.998 and the compatibility of the model Q- = 0.812 show that the proposed model is compatible with experimental results and the model is highly reliable. Optimal conditions for the hydrolysis process are 6.679 hours and enzyme concentration of 29.352U/ml at 60.015"C, resulting in the highest resistant starch of 6.04'^.
Keywords; Modification ofrice starch, optimized, resistant starch (RS),
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