Purification of Fructooligosaccharides Using Simulated Moving Bed Chromatography

Korean Chem. Eng. Res., Vol. 43, No. 6, December, 2005, pp. 715-721 Simulated Moving Bed 크로마토그래피를이용한프럭토올리고당의정제 j mp * l p, *r l 402-701 p}e n 253 (2005 9o 30p r, 2005 11o 24p }ˆ) Purification of Fructooligosaccharides Using Simulated Moving Bed Chromatography Nan-Suk Oh, Chong-Ho Lee* and Yoon-Mo Koo Department of Biological Engineering, *ERC for Advanced Bioseparation Technology, Inha University, 253, Yonghyun-dong, Nam-gu, Incheon 402-701, Korea (Received 30 September 2005; accepted 24 November 2005) k SMB Š p, v, Š m ( dšv, dšv)p t Š m p l o n m. SMB nr s p p rp e t l pp v k p (triangle theory)p rvž(standing wave) qpp. l pp e l m p p. Š m p nr t l p. pp q o r ˆl p l r m p l rn m. v rn dšv, dšvp p p p p v t vp Š m l l l m p v kk. Š m p p p m s l d p l. vl r } l ph sr p m l e p r eˆ p. h Abstract The SMB chromatography is used to obtain high purification of fructooligosaccharides (FOS), the mixture of kestose and nystose. SMB operation condition is usually determined by triangle theory or standing wave design when reactions do not occur within columns during experiment. Some of the reactions in columns may considerably affect experimental results. FOS can be hydrolyzed and converted into glucose and fructose during operation. To include the effect of reaction, the concentrations of each component at steady state after hydrolysis were used in simulation. The obtained simulation values are well matched with experimental results except sucrose. For sucrose, the experimental results were different from expected one due to the existence of an intermediate component. FOS is easily hydrolyzed and converted into glucose and fructose in more acidic condition and at higher temperature. Hydrolysis reaction can be prevented by the pretreatment of separation resin with NaOH as well as operation under lower temperature. Key words: Simulated Moving Bed, Fructooligosaccharides, Sucrose, Glucose, Hydrolysis 1. m p q 2~10 p, p rp r, d p pp,, r, rqqn p r p p. m p p p tep 1970 l v l, 1980 l Š m (fructooligosaccharides)p n eq m. Š m p, kž, kdž d, nl,,, ƒ p k}, p e l o p }l vpv k p. p o Aureobasidium sp[1, 2] To whom correspondence should be addressed. E-mail: ymkoo@inha.ac.kr Aspergillus niger[3] pn l vm p eˆ pp lrp p. Š m p vp β(2 1)p o l p l p vp p dšv(β-d-fru((2 1) 2 -α-d-glucopyranoside, GF 2 )), dšv(β-d-fru((2 1) 3 -α-d-glucopyranoside, GF 3 )), Š e- dšv(β-d-fru((2 1) 3 -α-d-glucopyranoside, GF 4 )) pp l l p p l GF n p ˆ [4]. Fig. 1l GF n p r sep ˆ l. Š m p l op rp pep n l p e ~ r f p. Š m p ol l vm pp vp l o rr rp e ~k. Š m 715

716 m ps o Fig. 2. Schematic explanation of simulated moving bed. Fig. 1. Chemical structure of fructooligosaccharides. n=1: 1-kestose, n=2: nystose, n=3: fructofuranosylnystose p rp n tn. p rp rr rl j p p Š f l n p + 2+ Na Ca ~ p kpm v rn pp p p e l ~ vp p n p. Š m p p phm p m l pl [5, 6]. p ml m k n CO 2 m p l k p p v n r t p s p p., rp p p p rn r l p m l nr. m p d s l p lv. p o vp r} rp p m l rp p lrk Š m p ep tp p. SMB p 1960 UOP l p k [7, 8], p pn o p-xylenel rk kp p v~ [22]m r l[9, 10, 21] k v[11, 12], v m Œ [13, 23] k kp v l. SMBp o pp o pr e p nvpp f r p p p p l rp p [14~16]. rm p k nvp vp k p p nvl extract p rm p nvp vp r p p nvl raffinate p. p l rp l l pp p p, extract, raffinate [17, 18]. SMB 4 p lp p m p o l lp tn l p [19]. rp 4 p lp l p SMB Fig. 2l ˆ l. SMB p l rp e rl, p p p lp p. SMB e p q v p, p rp pq e p m srp SMB e p. pp pv k l e t pp o43 o6 2005 12k pl m l. e l SMB pn l l rp Š m p q m. p e l rp k o l l vp p p p mp s p Š m p pl vp s p er m e m p m. p q o SMB e s extractm raffinatel lp e l r ˆl vp kp l r l l l e m k., SMB e s pp eˆ o v r } l ph t l sr m l. p p r p f pp mpp p m. 2. h m p ež p e ~ r pn m, v, dšv, dšv pp 45 Brix p. m p r} rp f ppm vm kpm v e v p p r m. p p 3 v n m v Finex l p p s dˆp l p rp mp pm(so 3 )p p kpm v f + Na ~ ˆp. SMB e p l 2 j, 8 p p (21.5 2.5 cm)p n mp Omni l p m. p p o extractm raffinate v o 4 p n lp FPLC 2 (amersham phamacia biotech), HPLC 2 (young lin instrument) pn l o p sr m. p p e p n o rp 3 v n m. v r } r p o 4ÍNaOH 3.5 ml/min(2 bed/hour)p p eˆ 3 v t p ph v kl tl., 65 Cm o 50 o C l m l m p m. s

l r o 12e k m p l pr o p q eˆ p p r m. p r rp p Kromasil 100-10 NH 2p(250 4.6 mm) HPLC p pn mp p p acetonitrile/water/v/v/75/25s p acetonitrilep HPLCnp J. T. Bakerl p m. l n Shimadzu p r p LC-6AD, SIL-10AD VP q tp, RID-10A r HPLC edšp. 3. 3-1. j y m o l tn vp, v, dšv, d Šv f r~p 21Í, 24Í, 36Í, 16Í p. p vp p r kk o r p p pn m. l kp 20Í, 40Í, 60Í, 80Í, 100Í 5 e mp Fig. 3l ˆ l. e p lp p (1) ep rn l p l p k r l kp mp p (2)ep pn l m [20]. Table 1l ˆ l. Š m p rr 717 Q i + 1 = Q i ( V F V 0 )( C i + 1 C i ) + ------------------------------------------------------ Va Q = ac (2) l Q r p vp k, C p l k p vp, V F ~, V 0 o~ p, V a r p, V c p p. 3-2. j er e p nr r e p m rp. nrp rs p o rvž p p pn m. rvž p p SMB r qpp o Ma and Wang[15]p m k m k l e l rn p. p p o nvp vp p l IVl, ˆ p l IIl p pm d nvp Table 1. Linear isotherm parameters of glucose, sucrose, kestose and nystose Glucose Sucrose Kestose Nystose a 0.4005 0.2372 0.1563 0.0848 R 2 0.9999 0.9979 0.9957 0.9881 (1) Fig. 3. Determination of isotherm by frontal test. (a) glucose, (b) sucrose, (c) kestose, (d) nystose. Korean Chem. Eng. Res., Vol. 43, No. 6, December, 2005

718 m ps o Fig. 4. Design for four zone SMB process : standing wave design. --- -- : nystose, -.-.-: kestose, -----: sucrose,...: glucose Table 2. Operation condition of SMB Feed Extract Raffinate Eluent Recycle Switching time 0.30 ml/min 1.68 ml/min 1.20 ml/min 2.59Gml/min 5.65 ml/min 8.52 min vp p l, ˆ p l IIIl plk l. l IIm III v p rp p lv l I IV v p mmp v o p p p mt. p ov o lp o nvp tn p v r p m p v k qp vr p m l ps qpl p. p e l vr p m p qpp rn m. e l n p, v, dšv, dšvp v. Š m p raffinate o l,, v extract o l l o l1p p ˆ, l 2 dšvp ˆ, l3p vp, l4 dšvp p p qp m. p ˆ p Fig. 4l ˆ l., p p p llv nrs p Table 2l e p s p e p m. ol l mp s p p l. pp m v kp p q o e s p l r l m. l r ep (3)e, Table 3 m e p Fig. 6l ˆ l. p v e rl m ll l rn l ˆ l. Table 3. Concentration of component in feed before and after Hydrolysis Before(mg/ml)) After(mg/ml) Glucose & fructose 128.97 345.57 Sucrose 120.39 78.15 FOS 300.6 126.22 o43 o6 2005 12k Fig. 5. Comparison of experimental and theoretical concentration profiles: without reaction at the end of a switch time. (a) ---- : simulation of sucrose, -----: simulation of glucose : experiment of sucrose, : experiment of glucose (b) ---- : simulation of kestose, -----: simulation of nystoseg : Gexperiment of kestose, : experiment of nystose C Feed *v Feed = C Extract *v Extract + C Raffinate *v Raffinate o el C, v o p ˆ. Fig. 5l p p, v kp p p m p p e p, vm Š m p l m p e p ll. pp v kp n l n p p Fig. 6 p rn l dšv, dšvp p m p p mp vp p p m. po v Š m l vp s vp p rp t vp l e m p v kk. 3-3. j m 3-3-1. php m Š m l m p pq t php (3)

Š m p rr 719 Fig. 7. The ph change in column. : without NaOH 65 o C, :Gwith NaOH, 65 o C, :Gwith NaOH, 50 o C. Fig. 6. Comparison of experimental and theoretical concentration profiles: with reaction at the end of a switch time. (a) --- - :Gsimulation of sucrose, -----: simulation of glucose : experiment of sucrose, : experiment of glucose (b) ---- : simulation of kestose, -----: simulation of nystose : experiment of kestose, : experiment of nystose [5, 6]. v r } l p php m php l r o e p m. e p 65 C m r v 40Í NaOH } o r p s l m p 12e k q eˆ 2e p e v m. p e p ph r HPLC l p pp r m. php r } r p pp Fig. 8(a), (b)l ˆ l. ph r } rp p l Š m p. p 12e r } r dšv p 5.3Í, 33Í, dšv 2.2Í, 12.4Í, v 42.1Í, 67.6Í mp p 347Í, 245Í v m. r } p f p ph t l ov pl p r eˆ pl. 3-3-2. m p m Š m p l m p npp m p [5, 6]. ol l p rp k 50 Brixp l p lv l p r v. r o 50 o C~65 Cp p m l o. m p kk o r } v pn l 50 Cm o 65 Cl m p 12e k q o eˆ 2e p e v l m. 12e 65 Cm o 50 Cp dšv p o 33Í, 88Í, dšv 12Í, 78Í m. v 65 Cl p 67Í o mv 50 Cl kp 114Í o v m. v v po Š m p p v j vp p p m l lp kp k v p., p 65 o C, 50 Cl kp o 245Í, 133Í v m. m l p p Fig. 8p (b), (c) p. 3-4. o m i mi Š m vp kp m p kp v mp pl p ep. vm p p l Š m p p p l v pp p p l pp r kk o e p m. e, Fig. 9m 10l p p v Š m p v kk. pp l pp pl v k l rp l pp (4~6)ep ˆ p. GF3 k 3 GF 2 +F (4) GF2 k 2 GF+F (5) Korean Chem. Eng. Res., Vol. 43, No. 6, December, 2005

720 m ps o Fig. 9. Hydrolysis of sucrose in columns as a function time. : glucose, : fructose, : sucrose, : total concentration Fig. 10. Changes of concentrations in columns as a function time. : glucose, : fructose Fig. 8. Hydrolysis of fructooligosaccharides in columns as a function time. :Gglucose and fructose, :Gsucrose, :Gkestose, :Gnystose (a): with NaOH, 65 o C (b): without NaOH, 65 o C (c): with NaOH, 50 o C GF k 1 G+F (6) G:, F:, GF: v, GF 2 : dšv, GF 3 : dšv o43 o6 2005 12k 4. Š m p on vp l l p p rr rp n tn. q Š m p rr Š p pn pp p e l l rp rp SMB rn m. v f rl n p mp ~ p kpm vm p p p n p ˆl r p r l ml nr. p s l pl v s p rm. SMBp r nrs p r o qpp p p pv kk pp pl m l. SMB n vp l rp o p pp ˆ k pp t ˆl p lv Š m p vp l rp pp n

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