cau1 chung giong va quy trinh san xuat enzym amylaza tu nam moc I.

T ng quan v enzym amylase

1. Amylase l m t h enzyme r t ph bi n trong th gi i sinh v t . Cc enzyme ny thu c nhm enzyme th y phn, xc tc phn gi i lin k t n i phn t trong nhm polysaccharide v i s tham gia c a n c: R.R + H-OH RH + R OH 2. C 6 lo i enzyme c x p vo 2 nhm: Endoamylase ( enzyme n i bo ) v exoamylase ( enzyme ngo i bo ). - Endoamylase g m c -amylase v nhm enzyme kh nhnh. + Nhm enzyme kh nhnh ny c chia thnh 2 lo i: Kh tr c ti p l Pullulanase ( hay -dextrin 6 glucosidase); kh gin ti p l Transglucosylase (hay oligo-1,6-glucosidase) v maylo-1,6-glucosidase. Cc enzyme ny th y phn cc lin k t bn trong c a chu i polysaccharide. - Exoamylase g m c -amylase v -amylase. y l nh ng enzyme th y phn tinh b t t u khng kh c a chu i polysaccharide . 3. C ch t tc d ng c a amylase l tinh b t v glycogen

III. Gi i thi u v ch ng n m m c Aspergillus oryzae

Asp. oryzae l m t lo i n m vi th thu c b Plectascales, l p Ascomycetes ( nang khu n ). C th sinh tr ng c a n l m t h s i bao g m nh ng s i r t m nh, chi u ngang 5-7 m, phn nhnh r t nhi u v c vch ngang , chia s i thnh nhi u bao t bo ( n m a bo ). T nh ng s i n m ngang ny hnh thnh nh ng s i ng th ng g i l cu ng nh bo t , c c quan sinh s n v tnh. Cu ng nh bo t c a Asp.oryzae th ng di 1-2 mm nn c th nhn th y b ng m t th ng. Pha u cu ng nh bo t ph ng ln g i l b ng. T b ng ny phn chia thnh nh ng t bo nh , thun, di, g i l nh ng t bo hnh chai. u cc t bo hnh chai phn chia thnh nh ng bo t nh vo nhau, nn g i l nh bo t . nh bo t c a Asp.oryzae c mu vng l c hay mu vng hoa cau c i m c a gi ng Asp.oryzae giu cc enzyme th y phn n i bo v ngo i bo ( amylase, protease, pectinasa, ), ta r t hay g p chng cc kho nguyn li u, trong cc thng ch a ng b t, g o h t nh ng khng c r a s ch, c n b bia, b r u, l i ng, b s n Chng m c v pht tri n c khi thnh l p m c, c mu s c en ,vng Mu do cc bo t gi c mu s c. Cc bo t ny, d b gi cu n bay xa v r i vo u khi g p i u ki n thu n l i s m c thnh m c m i.

III. Quy trnh s n xu t

Ch ng m c gi ng: Aspergyllus oryzae B o qu n gi ng: Cho ct vo ng nghi m, ti t trng trong autoclave( n i h p kh trng) 130oC trong 30 pht. Tr n bo t vo ct. S y chn khng < 40 oC ( m 5%). Dng parafin r n un ch y v ln nt bng. Th i gian b o qu n: 1 n m ho c h n. Nguyn li u: * Ngu n tinh b t: cm g o: ch a kho ng 20% tinh b t, 10 15% ch t bo, 1014% protein, 8-16% cellulose, cc ch t ho tan khng ch a nit 37-59%. * Cm khng c ch a hm l ng tinh b t d i 20-30%, khng c v chua hay ng, khng hi mi m c, m c a cm khng qu 15%, t p ch t c khng qu 0,05%. * Cc ph ph m thm vo t ng thong kh cho mi tr ng nui c y: tr u, m t c a

Quy trnh
+ X l nguyn li u:cm g o, tr u v m t c a c x l Tr u cho vo v i t l 20-25% so v i kh i l ng cm g o. lo i b cc t p ch t.

+ Thanh trng: Nhi t thanh trng l 95 oC trong 60 pht. ng th i ph i thanh trng khay ln men. + R i nguyn li u ln khay c kch th c 2x3 m, dy 2-3 cm, dng 3kg mi tr ng/khay. + Sau khi lm ngu i, ti n hnh c y m c gi ng v i n ng 5-10% so v i kh i l ng c a mi tr ng m i khay. + a khay ln cc gi + Nhi t phng nui c y c gi 25-30 oC, m 60-65%, ng th i ph i thong kh.

n kho ng 30-32 gi sau khi c y gi ng, ta thu nh n enzyme th. C n thu nh n enzyme th vo th i i m tr c khi n m sinh bo t v khi n m sinh bo t l lc qu trnh t ng h p enzyme a y u i. Do c n m t b c nui c y th nghi m xc nh th i i m thu nh n enzyme th. Ch ph m enzyme th thu c ch a t bo n m, c ch t, n c v enzyme

gi ho t tnh c a enzyme, s y ch ph m th ny nhi t 30-40 oC, m sau s y l 10%. + Nghi n m n: ch ph m enzyme th c nghi n v i ct v b t th ch anh. Ct v b t th ch anh c r a s ch, s y kh tr c. + Trch ly: sau khi nghi n, dng n c trch ly. C 1l enzym th, cho 4-5l n c, khu y nh v sau l c l y d ch. + L c: d ch sau trch ly l c b ng l c ti p tuy n. + K t t a enzyme: k t t a c th c hi n b ng ethanol ph i lm l nh c dung d ch enzym th v c nh ng tc nhn k t t a trnh lm m t ho t tnh enzym.Khi ch t lm k t t a enzym vo dung d ch enzym th ph i h ts ct t trnh hi n t ng bi n tnh. Nhi t t 3-10 oC v dng 2 l ethanol cho 1 l enzyme. + Enzyme thu c l 1 h n h p nhi u enzyme khc nhau nh : amylase, proteinase, cellulase Do , ph i ti n hnh s c k l c gel thu c enzyme amylase. Gel s d ng l Sephadex G200. + S y: s n ph m sau s c k c s y nhi t 30-40 oC b o m ho t tnh cho enzyme, m s n ph m l 5%. +

quy trnh
Nguyn li u X l Thanh trng Lm ngui Thnh ph m Bao gi S y S c k thu nhn enzym th K t t a enzym L c Trch ly

Gi ng VSV

Ph i tr n Nui c y Thu nh n enzym th Nghi n m n