CVv400S22013112.pdf

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Tgp chi Khoa hoc - Cong nghe Thuy sdn So 2/20U

KET QUA NGHIEN COU DAO TAO SAU DAI HOC

SANG LOC CHIJNG VI KHUAN CO KHA NANG SINH GELATINASE Tlf DAT \'k XAC DLSH MOT S6 TINH CHAT CUA DICH ENZYME THO

SCREENING OF A GELATINASE PRODUCING BACTERIUM FROM SOIL AND CHARACTERIZATION OF THE CRUDE ENZYME EXTRACT

Tran Qudc Dam', Nguyin Thi Thanh HdF, Nguyen Anh Tuan', Nguyen Minh Tri' Ngaynhanbai. 21/8/2012; Ngay phan bien thong qua. 25/01/2013; Ngay duyet ding: 15/5/2013

T O M TAT

Gelatinase co khd ndng thuy phdn gelatine thdnh cdc peptid vd acid amin hay con goi Id gelatin thuy phdn dupc i^

dung trong nhieu ngdnh cong nghi^. Muc tieu cua nghien aiu ndy Id nhdm thu nhgn gelatinase tir cdc vi khudn trong iai vd khdo sdt mot s6 ddc dnh cua enzyme ndy. Chung tdi dd tien hdnh phdn lap vd tuyen chon chung vi khudn co kha nong sinh gelatinase tren moi truang ddc trunggelatine-agar (gelatine 1,5%; agar 1,5%, p^ton 0,4%, cao ndm men 0,1% va pH 7.2). Ket qud tuyin chpn duac chimg Bacillus sp. C7 cd khd ndng tdng hgp gelatinase cao. Enzyme gelatinase thu duqc hogt dong toi uu a nhi^t do 50 °C, pH 7,5. Cdc ion Mr^* vd Fe^' ldm tdng hoat dnh aia enzyme vd ian Ca'^ iam gidm hogt dnh gelatinase. Nghien cuu ndy se Id ca sa cho cdc nghien ctiu dip theo ve thu niton gelatinase tinh khiet tir chung BacUba sp. C7 vd ung dung enzyme ndy vdo ddi sdng sdn xudt

Tit khoa: Gelatin. Gelatinase, Bacillus ABSTRACT

Gelatinase can hydrolyze gelatine into peptides and amino acids which is called hydrolyzed gelatin and this is useiiiii industries. The present research aims to obtain gelatinase from bacteria in soil and define some properties ofthe gelatinast Gelatinase producing bacteria were isolated and screened on the selected gelatine-agar medium (1 5%geladn, 1 5% agar.

0.4%peptone, 0.1% yeast extract and pH 7.2). The result of the screening has indicated a strain Bacillus sp. C7 of wA high gelatinase production activity. The optimal pM and temperature ofthe gelatinase from Ihe strain Bacillus sp. C7 HCTF found to be at 7.5 and 50°C. respectively. Ion Mn^" and Fe^' were shown to increase the gelatinase activity whereas iha activity was reduced by ion Ca^'. The study is the basis forfunher researches on obtaining purified gelatinase from siraai Bacillus sp. C7 and application of this enzyme.

Keywords: Gelatine, Gelatinase. Bacillus

I. DAT V A N D £ Djch thuy phan gelatin cd chira mgt Itrpng \tm (A;

Gelatn Id n>it polymer smh hpe ed nguon gic tir acid amin nhu glycine, proline, hydroxyprofine, collagen, la thdnh phin chu yeu eiia da, xuang va lysine, hydraxylysine... vd chiing de ding dupe hip md llm ket dgng vgt [7]. Gelatin duoc thiiy phan tgo thy vdo ca the. Day la cde thinh phin chire ning thdnh cdc peptid vd add amin goi Id djch thuy phdn quan trang. nd duoc irng dung trong cong nghidp my gelatin. Khi nang hda lan, kha ndng tgo nhO, tao bot phim vd diKrc pham de san xudt cic sdn phim ho hay hogt tinh ctiong oxy hda... la nhung Unh chit trp d u true collagen trong da vd syn [11].

quan trpng cua djch thiiy phan gelatin dupc img dung Qua trinh thuy phdn gelatin co the dinj":

rgngrailrongnginheongnghiipthucphlm[11.12]. thuc hign bing phuang phap hda hgc hodc t)ing

' Tran Quoc Dam' Ldp Cao hpe Cdng ngh^ Sau thu hoach 2010 - Trudng Ogi hoc Nha Trang

' ThS. NguySn Jh\ Thanh Hdi. ^ S Nguyen Anh Tuin, TS. ^Nguyin Minh Tri; Khoa Cong nghg thye phim - Tnrdng Ogi hpc Nha Trang

112 • TRUONG DAI HOC NHA TRANG

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Tgp chi Khoa hoc - Cong nghe Thuy sdn So 2/2013 phuang phap enzyme. Chung ta da bill, enzyme

ed cudng lyc xue tac mgnh ban r i t nhieu so vdi cac chat xiic tdc v i ca vd hiru eo khde, dong thdi nd cd the thye hign hoat dgng xue t i c d dieu kign dp suit vd nhigt do thudng [1. 2]. Tir nh&ng dgc diem tren cho thiy phuang phap enzyme gdp phin ndng cao chat luang sdn phim vd gidm thieu t i c dgng g i y o nhiem mdi Irudng Iir qua trinh sdn xult so vdl phuang phdp hda hgc. Vigc nghien cdu vd ung dyng enzyme cd y nghTa to Idn v i mat ly thuylt eung nhu ve thye te ap dyng [1]

Gelatinase la m i t enzyme thiiy phdn protein, ed t h i thiiy phdn gelatin thdnh cdc peptid vd cdc acid amin. Chiing ta cd the thu nhgn gelatinase tir nhilu nguin khdc nhau, trong dd Ihu nhin gelatinase Iir vi sinh vgt dupc chu y nhieu ban ed. Trong ty nhien e i nhieu ehiing vi khuin cd kha nang sinh ting hpp gelatinase nhu: Bacillus, Flavobactetium, Proteus.

Pseudomonas, Salmonella, Sarcina, Serratia, Shigella. Vibrio... [5, 6, 10] Vigc tuyin chgn chiing an todn, thudng duac quan lam nham dng dyng trong ngdnh cing nghe Ihyc phim [8, 9].

Hogt dpng cua enzyme phg thugc nhieu v i o dilu kiin mdi trudng md n i xue tde phdn irng [3].

Vige xie d|nh dieu kien hogt dgng xue t i c thieh hpp cQa gelatinase giiip cho vigc dng dyng thiiy phin gelatin bing gelatinase m i l cdch hi^u qua.

Myc tieu cua nghien eiru nay id sdng igc dupe ehung vi khuan cd khd ning sinh l i n g hpp gelatinase ed boat tfnh cao vd khdo sdt mpl s i dieu kiin hogt dgng ciia enzyme ndy

II. VAT Lieu VA PHUaNG P H A P N G H I D N CO'U 1. M i l tru-d-ng phdn l | p vd nudi sinh enzyme

M i l tnj'dng phan ldp (MT1)' sir dyng mdi Irudng thgch gelabn-agar cOa Smith v i Goodner cd thinh phin nhu sau' gelatin 1,5%, agar 1,5%, pepton 0,4%. eao nim men 0,1%. Mdi tnrdng dupc dieu ehinh ve pH 7-7,2 v i v i tning d 121 "C trong 15 philt trudc khi su- dyng [6].

M i l trudng nudi sinh enzyme gelatinase (MT2):

su dyng mdi trudng phan ldp khing b i sung Ihinh phin agar.

2. M l u d l t phan lap vl khuan

M l u d l t dupe lly tgi Thdp Bd, VTnh Phudc, Nha Trang, Khinh Hia. Qud Irinh chuin bj mau nhu sau. Da ed tra dupc Ci trong d l t cho den khi bj mun ra. Khi da cd bj phdn buy, chuin bj 100 mi dung djch gelatin 1%, d i xuong phin d l t da i da ca, dinh ky

3 ngay mdt i l n , lien hdnh lien tuc Irong khoang 2 tudn. Sau dd. lly 1 g m l u d l t d do sdu Iir 3 - 6 cm cho vao 10 ml nude v i InJng, trin d i u vd de on djnh trudc khi phdn l|p.

3. Phan lap chung vl khuin sinh gelatinase M i u dupc phin lap tren mdi trudng MT1 bang ky thudt cay ria, u d 3 7 1 Irong 48 gid. Cac vi khuin cd khd nang phdn cat gelatin se tao vdng trong xung quanh khuan Igc. Chgn cac Wiuin lac cd hinh thai khie nhau dem nudi cay v i giu chiing [4, 6].

4. Tuyin chon chung vi khuan sinh gelatinase cao n h i t

Tien hdnh nuii cly cdc chiing bdn citng mgt dTa thach chira m i l tnjdng MT1. u d 37''C. Cdc chiing vi khuan phdt trien tao vdng phan giii xung quanh khuin lae. Tien hanh do dudng kinh khuin lac v i dudng kinh vdng phan giii tgo ra xung quanh chiing VI khuan dd. Chiing ed ty ie dudng kinh vdng phan giai/dudng kinh khuan Ige Idn nhit Id ehOng ed khi nang sinh gelatinase eao nhit trong nhdm ehiing phan lap dupc [4]

5. Thu djch gelatinase tho

Chung tuyen ehpn 6ir(yc nuii eay Irong mdi trudng nuii sinh gelatinase MT2 d nhigt dp phong, trin may lie 180 vong/phOt trong 48 gid. Canh trudng nudi cly dupc ly Idm 4000 ving/phiit trong 15 phut, d 4''C d i logi bo sinh khii vi khuin. Phan d[ch cdn iai ehira gelabnase va m i t s i chat hda tan khac ggi la dtch gelatinase thd.

6. Xac djnh hogt dp cua gelatinase Hogt do enzyme gelatinase dupc xdc djnh dya vdo lupng mg nito amin hinh thdnh trong qui Irinh thiiy phdn gelatin. Mgt dan vj hoat do gelatinase la luang enzyme thiiy phin gelatin tao thinh 1 mg nita amin Irong 1 gid d 40''C va pH thich hpp [1].

Tien hanh IhCiy phdn gelatin 1%, ty Ig E/S la 1/2, pH 7. d nhiet dd 40''C Irong 1 gid de xde djnh hogt do djch gelatinase thd.

7. 0jnh lupng nito- amin

Hdm luang nito amin duac xie djnh dya vao him lupng nita amin-amoniae (X,) v i hdm lupng nita amoniac (X^^^j).

Dam nita amin (X^) dupc tinh nhu sau' x..= x,-x„„,

- Ham lirgng nita amin-amoniac di^ac xac djnh bang phuong phap formoi (TCVN 3707 - 90)

- Ham lu'p'ng mto amoniac duoc x^c dinh bing phuong phSp Chung cat (TCVN 2626-98).

TRi;dNG OAi HOC NHA TRANG ' U S

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Top chi Khoa hoc - Cong nshe Thuy s So 2/2011

8. Bo tri thi nghiem xac djnh mot so tinh chSt cua djch gelatinase tho Dich gelatinase tii6

3

Thuy phan gelatin 6 cac

dieu kien khac nhau

)

-Nhilt do: 30-70oC - p H : 5 - 9

- Ion kim loai: EDTA, Cu2-l-, Mg2+, Mn2+.

Fe2+ va Ca2+

31

Dieu kien thich hgp

Hinh 1. Sff do bo tri thi nghifm xac djnh 4nh hudng cua mOt so yeu t6 den hoat dg gelatinase tir chiing nghiSn cihi

D e x d c dinh m i l s o t i n h e h i t cOa d i c h g e l a t i n a s e t h i , p h a n i r n g t h u y p h d n d j c h g e l a t i n b i n g enzyme niy d u p e t h u c hign i l n l u p e d e i e d i e u k i i n nhiet d g , p H v d ion k i m logi k h i c n h a u . T i e n h i n h t h i n g h i g m tirng yeu t i d o n v d l c d e g i d I n d a e h p n v d e i e y i u t o c d n Igi gitr n g u y d n k h d n g dot t h e o b o t r i I r o n g h i n h 1 . 9 . D i n h d a n h v i k h u a n

C h i i n g v i k h u i n d u a c djnh d a n h b i n g p h u o n g p h d p gidi t r i n h t y g e n 1 6 S r O N A v d s o s i n h v d i ngan hSng g e n t r e n N C B I b i n g edeh s i r d y n g c d n g e y B L A S T (http://nebi.nlm n i h . g o v / ) .

M l u vi k h u i n C 7 d u p e g i r i (^nh d a n h tai C d n g t y T N H H N a m K h o a - T h d n h p h i H o C h i M i n h (htlp://www.

n k - b i o t e k . e o m . v n / ) .

III. KtJ Q U A N G H i e N CO'U V A T H A O L U A N

1 . P h d n l d p v a t u y e n c h o n c h i i n g v l k h u a n s i n h g e l a t i n a s e e a o 1.1. PhSn t$p cac chung vi khuan tiF dk sinh tong hQp gelatinase

S a u 4 8 g i d c l y m a u p h a n lgp t r e n m i i I r u d n g M T 1 I h i I h l y c d n h i e u k h u i n l a c p h i t t r i e n t a c h b i i t nhau. Ket q u i p h d n l g p d u p c 1 2 e h u n g vi k h u a n c d k h a n a n g sinh g e l a t i n a s e ,

1.2. Ki4m tra kha nSng sinh gelatinase cua c^c chung vi khuan phan l$p dut?c V i i c k i e m tra k h a n a n g sinh g e l a t i n a s e cCia e a c e h i i n g v i k h u i n p h a n IdP d u p c Id c a s d c h o v i g c t u y e n c h g n e h u n g t h i c h h p p d e t h u n h d n g e l a t i n a s e . K i t q u d nuoi e l y edc c h i i n g p h d n lap d u p c t r e n m i i t r u d n g M T 1 c h o t h a y c d e c h i i n g d i u p h i t t n l n v d l a o v d n g p h d n giai x u n g q u a n h s i n h k h i i c u a e h u n g D i e u n d y c h o t h i y c d e c h i i n g vi k h u i n p h d n l i p d u p c d i i u e d k h i n a n g s i n h g e l a t i n a s e . K e l q u d b u d e d i u c d n c h o thay eae e h u n g vi k h u a n C l . C 2 , C 3 . C 4 , C 5 v d C 7 l i n h d m c h u n g c d k h i n i n g sinh g e l a t i n a s e c a o I r o n g 12 c h i i n g p h a n l d p d u p e ( h i n h 2 ) .

1.3. Tuy4n chon chung vi khuin sinh gelatinase cao nhit

De Ihu nhgn gelatinase cd hogt tinh cao tir c i c chiing vi khuin phdn lgp dupc thi vi^e luyen chgn chung phii hpp la yeu to quyet dmh. Chiing phu hpp id chitng cd kha ning sinh gelatinase eao vd thugc nhdm GRAS. Sau 72 gid

nudi ely cdc chung C l , C2, C3, C4, C5 vd C7 tren m i l trudng MT1 thi t h i y eae ehiing vi khuin deu phdt tnen vd tgo vdng phdn giai Idn. Ket q u i do dudng kinh vdng phdn giai vd dudng kinh khuin Igc cua chung vi khuan tuang ung dupc Irinh bdy trong bing 1. Chiing vi khuan C7 la ehung ed k h i nang sinh gelatinase eao nhat trong nhdm chiing tuyen chpn.

Bang 1. D u v n g kinh vdng phdn giai vd d u ^ n g kinh k h u i n lac cua cac chiing vi k h u i n t r i n m i i tnrimg MT1 sau 72 gid- nuoi cay

Hinh 2. Vong phin giiti gdattne cua cic chiing vi khuan phiD Ifp

Chung vi khuan C1 C2 C3 C4 C5 07

OiKrnq kinh v6nq phan qi^i (P) mm 22,S 2 1 , 5 11 1 0 , 5 12,5 20

S i r i n g tcinh khuSn lac (d) nim 1 2 , 5

8 5,5 4 , 2 5 5 5.5

Ty 1$ Old 1,80 2,69 2,00 2 , 4 7 _ _ 2,50 3,64

1 1 4 • T R U O N G O A I H O C N H A T R A N G

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Tap chi Khoa hoc - Cong nghe Thuy san So 2/2013 2. Dinh danh chijng vi l<hu^n C7

Ket qua giai trinh tu doan gen ma hoa 16S rRNA dai 472 bp tren may CEO 8000 duoc trinh bay trong hinh 3. Cac trinh tir gen n^y duoc so chu5i b§ng chuang trinh BLAST SEARCH tren ngan hang du' lieu gen NCBI dugc trinh bay trong bang 2.

GTACCGCCCTATTCGAACGGTACTTGTTCTTCCCTAACAACAGAGCTTTACGATC CGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCG GAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGT GTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTTGCCTTGGTGAGCCGT TACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCCGAAGC CACCTTTTATGTTTGAACCATGCGGTTCAAACAACCATCCGGTATTAGCCCCG GTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTC CGCCGCTAACATCAGQGAGCAAGCTCCCATCTGTCCGCTCGACTTGCATGTATTAG GCACGCCGCCAGCGTTCGTCCTGAGCCAGGATCAA

Hinh 3. l > i n h fu- gen 16S rDNA cua chung vi khuiin C7

Bang 2, Ket quii tra cipu trinh tir gen 168 rDNA cua chijng 07 tren BLAST SEARCH A c c a i o n

C P 0 0 2 9 0 6 . 1

J N 3 9 9 2 1 9 . 1

J F 7 4 9 2 7 8 . 1

K C 2 5 0 1 2 3 . 1

C P 0 0 3 6 9 5 . 1

D Q 3 4 5 2 8 9

N R 0 2 4 6 9 3

Description B a c i l l u s subtilis s u b s p . subtilis R O - N N - 1 . c o m p l e t e g e n o m e .

B a c i l l u s s u b t i l i s s t r a i n D X ' 3 1 6 S n b o s o m a i R N A g e n e , partial s e q u e n c e .

B a c i l l u s subtilis s t r a i n A T C C 6 0 5 1 1 6 3 n b o s o m a i R N A g e n e , partial s e q u e n c e .

B a c i l l u s subtilis s t r a i n B A B - 1 1 5 1 6 S n b o s o m a i R N A g e n e , partial s e q u e n c e

B a c i l l u s s u b t i l i s s u b s p subtilis s t r B S P 1 , c o m p l e t e g e n o m e .

B a c i l l u s l i c h e n i f o r m i s 16S n b o s o m a i R N A g e n e , partial s e q u e n c e .

B a c i l l u s m o j a v e n s i s streiin I F 0 1 5 7 1 8 1 6 S ribosomal R N A , partial s e q u e n c e

Max

8 7 2

8 5 9 Total

8 7 2

8 5 9 coverage

1 0 0 %

9 9 % Expect

0.0

0.0 Max ident 1 0 0 %

1 0 0 %

9 9 . 8 %

Tir ket qua tra cuu Irinh l y gen 16S rDNA Iren BLAST SEARCH eho Ihly, chung C7 tuang ding 100% vdi S. subtilis vd ding thdi cung tuang ding 100% vdi e. lichenKormis va 99,8% vdi B. mojavensis (bing 2). Dieu n i y cho phep k i t lugn chung C7 thuic chi Bacillus.

3. Mpt so tinh chat cua enzyme thu du-g^;

Phdn irng do enzyme xiic t i e phy thudc vdo nhieu yeu t i nhu nong d i enzyme, b i n chit va ning dg ca chll. nhiet dd vd pH cua mdi trudng, cdc ion kim loai vd ede chit khde.. Trong nghien cuu niy, chting t i i t i i n hdnh xdc djnh pH, nhigt d i thieh hpp cOa enzyme thu dupc khi thuy phin gelatin vd anh hudng cCia mpt so ion kim loai den hogt tinh cda enzyme.

3.1. Anh ftmSng cOa nhi^t dd d4n hoat tinh gelatinase Nhigt dp phan irng anh hudng mgnh me den hoat linh xiic t i e cua enzyme, mdi enzyme cd m i l

nhigt do phan ung thieh hpp. Trong gidi hgn nhiet d i chua Idm biin linh enzyme, hoat ti'nh enzyme ting khi nhigt dd ting. Tuy nhien, khi nhiet do ldng den mdt gidi hgn nao dd thi hogt tinh ciia enzyme Igi gidm [1]. De khdo sdt nhiet do t i i uu cua gelatinase tip Chung Bacillus sp. C7. phin irng dupc thye hien d cie nhiet dp khie nhau Iir 30+70°C.

Hinh 3. Anh huong ciia nhiet dp phan irng den hoat tinh cua gelatinase tu chung vi khuan C7

TRL/ONG OAI HOC NHA TRANG - U S

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Tgp chi Khoa hoc - Cong nghe Thuy sdn So 2/2013 Ket q u i khio sdt eho thiy nhigt dp phan ung thich hpp cua gelatinase t u chiing Bacillus sp. C7 Id 50»c, ddy la enzyme ua nhigt va c i khoang nhidt do boat dgng tuang d i i ring tfr 30 + 60°C (hinh 3). Nhu vgy.

gelatinase e i the ung dyng vdo eac qua trinh ed gia nhigt nhung khing qud cao. 6' nhigl do la 50°C, khi t>ng dyng gellainase v i o qud trinh s i n xult cd the ban c h i dupe sy phdt trien cua m i l s i vi khuin cd the lim giam chit lupng sdn phim.

3.2. Anh hi/ang cua pH den hoat tinh cua gelatinase 0.40.

Hinh 4. Anh buong cua p H phan ung d i n boat tinb gelatinase tir chiing vi khuan C7

Gia tn pH mdi trudng ed anh hudng Idn den toe do phan irng eua enzyme, mdi enzyme chl hogt ddng tit tai mpl hi s i pH xde djnh gpi Id pH hogt dgng lit thieh cOa enzyme [1.12]. Tiiy thuic vdo bdn chll enzyme ma pH enzyme hogt ding thich hap de enzyme boat dgng ed the tmng tinh, kiem hoge acid {1]. De tim ra khodng pH t i l uu eho hogt dgng eiia gelatinase tir chung Bacillus sp, C7 phdn irng gida enzyme ca chit dupe ttiuc hi^n d nhigt dp 50°C vd gid tn pH Ihay doi trong khoang ttr 5*9. Ket qua nghien euu cho thiy gelatinase tir chiJng Bacillus sp. C7 hogt ddng tot nhit trong khoang pH 7.0-^7,5, thugc logi enzyme ua pH trung tinh.

3.3. Anh hwdng cua mot so Ion kim loai den hoat tinh gelatinase

€X; BDfA CU2+ Wi2*- tJ^+ FeZ+ Ca2+ lOn kim logl Hinh 5. Anh hiromg ciia mgt so ion kim Ioai den bo^l tinh gelatinase tii chiing vi khuan C7 Cdc ion kim logi c i the kim ham hogc hoat hda su

hogt dpng cua cdc enzyme [1. 12]. De khao sit anh hudng cua ion kim loai den hoat tinh ciia gelatinase tir ehiing Bacillus sp. C7. djch enzyme duac xu ly ii vdi cic ion kim logi Cu2+, Mn2+, Mg2+, Fe2+, Ca2+

d nong dd 2 mM vd EDTA d nong d i 2 mM tiirdc khi xie dinh hogt do Ket qud nghien ciru tir hinh 5 cho Ihly, tinh chit vd mi>c dp anh hudng cua cie ion kim ioai den hoat linh gelatinase tir ehung Bacillus sp.C7

Id khae nhau. Khi xir 1^ gelatinase vdi EDTA thi ho?t tinh ciia enzyme giim mgnh, dieu niy cho thay gelatinase thuic nhdm enzyme phu ttiuic kim io?i (metailoenzyme). Kit qui cung cho thay hai ion kim logi Id Mn2+ vd Fe2+ Id hai y i u l i hoat hoa cua gelatinase Idm tdng hogt linh enzyme. Ngupc igi, i""

Ca2+ la yeu t i kim hdm Idm giam boat tinh gelatnase nen d n dupc loai bd. Trong khi dd hat ion Cu2+ va Mg2+ lim ttiay dii hogt tinh enzyme khong ddng ke.

1 1 6 - TRUONG DAI HOC NHA TRANG

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Tgp chi Khoa hoc - Cong nghe Thuy sdn So 2/2013

IV. KtJ LUAN v A KI^N NGHj 1. K i t ludn

- Phan lap vd tuyen ehpn dupe chung 07 cd khi nang sinh long hpp gelatinase cao n h l l trong so cdc Chung phan lgp dupc tren mdi tnrdng thgch gelabn-agar.

- Ket qua djnh danh bdng gial trinh t\i doan gen 16S rDNA dai 472 bp cho thiy chiing C7 ttiudc chl Bacillus, cd dp brang dong 100% vdi Bacillus subttlis, 100% vdi Bacillus licheniformis va 99,8%

vdi Bacillus mojavensis.

- Xac djnh dupc mit s i yeu to anh hudng den hogt dpng xue t i e phdn dng thuy phan eua gelatinase tir ehiing Bacillus sp. C7: Nhiet dp thich

hpp Id 50oC, pH thich hpp 7,5, c i c ion Mn2+ vd Fe2+ Id c i c yeu t i boat hoa vd Ca2+ Id cde yeu t i kim ham hogt tinh gelatinase.

2. Kien nghj

-Tienhinhgidi trinh ty doan gen 16S rDNA d i l ban hoae thir nghigm phdn ung hda smh d i dmh danh chinh x i e ehiing 07 d i n lodi

- Nghien cuu tinh che gelatinase tir chiing 07 va dp dung enzyme ndy nhim thiiy phdn gelatin

- Nghien ciru b i sung gelabnase vdo chupp s i n xult nude ndm nhdm t i n g cudng k h i nang phdn c i t cdc thdnh phan collagen tir da vd sun ed Idm t i n g h i m lupng dam acid amin cho sdn phim.

TAI LIEU THAM KHAO Tiing Viet

1 Nguyin Trpng Can, Nguyin Thi HiSn, Do Thi Giang, TrSn Thi Luyen, 1998. Cong nghe enzyme. NXB Nong nghi?pTP H6 Chi Minh.

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