Nghiên ứu ông nghệ xử lý nướ thải dệt nhuộm bằng enzyme laase

florida sau 24h.

LACCASE

C u trúc phân t c a laccase

               benzenethiol [ ]56          - dimethoxy-4-hydroxybenzaldehyde azine (syringaldazine, SYR), 1-naphtol, p-cresol (1-hydroxy-4-methylbenzene), 2,6-dimethoxyphenol (DMP) và 2-methoxyphenol

các hydroquinone là 2-methoxy-1,4-benzohydroquinone và 2,6-dimethoxy-1,4-          

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         3-diazenylbenzenesulfonic và 4-methylimino- cyclohexa-2,5-dienone 3 - axit diazenyl-benezenesulfonic m     

 1,2-Các  quá trình oxy hóa   trùng  

Laccase hong t i thích trong kho ng pH 4    i vt phenolic Khi

c vùng ki m thì ho t tính c a laccase b gi m, nguyên      nhân do anion nh      c ch laccase M       gim th oxy hóa kh c  t phenolic d b oxy hóa   by, ho t tính laccase các pH khác nhau là k t qu c a hai tác      d i l p c a pH là s   ch th oxy hóa kh laccase    t và tác d ng c ch    tru  ng ba nguyên t c    i v   t không ph     n       n s v n   chuy    i thích n m trong kho ng 2     c l i, tính b n c a laccase cao nht n trong kho ng pH ki m n m trong kho ng 8        b n nhit c, ph thuc vào ngu n g c c a vi sinh v t Nhìn chung,     laccase b n 30 50   o C và nhanh chóng m t ho t tính nhi    trên 60 o C Laccase b n nhi t nh  c phân l p ch y u t các loài thu c prokaryotae, ví d      thi gian bán h y c a laccase c  a Streptomyces lavendulae là 100 phút 70 o C và c a protein cotA  t loài  Bacillus subtilis là 112 phút 80 o C Th i gian bán h y c a laccase có ngu n     g c n ng là nh    70 o C vi 10 phút 80 o C [34 37] ;

Ficus racemosa L (cây sung), Weeping willowNicotiana tabacum    Prunus persica        

Pleurotus ostreatus, Phlebia radiate, Coriolus (Trametes, Polyporus) versicolor  Ascomycetes, Deuteromycetes [32]

Tribolium castanem, Manduca sexta, N cincticeps [ ]26 

V i ph r  t, laccase có kh   d ng các lo i thu c nhu m Tuy nhiên, m t s màu không th b oxy hóa ho c ch b           

28 oxy hóa m t ph n b i laccase do chúng có c u trúc quá c ng k nh ho c th oxy hóa         kh quá ca có th g n k t v i trung tâm ho    ng c a enzyme 

Theo k t qu phân tích LC MS, laccase t    Polyporus sp S113 có kh   h y màu RBBR thành hai s n ph m nh là sodium 1 amino 9, 10 dioxo 9, 10         dihydroanthracene 2  sulfonate và sodium 2  (3  aminophenyl) sulfonyl) ethyl sullfat Tuy khng phân t c a nh ng s n ph m phân h y nh        c tính không ít u Vì v y hi u qu   loi màu có th   lý thuc nhu m v n c  c ti p t c nghiên c u    n khi lo i b   c hoàn toàn các s n ph m trung gian nguy h i v i s c kh     i

Campos và cs [19] T hirsuta (THL1, THL2) và Sclerotium rolfsii       

   acetosyringone, 1-hydroxybenzotriazole (HOBT) và 4-hydroxybenzenesulfonic acid

Schizophyllum commune, N crassa, Polyporus , S rolfsii, T villosa Mycelioph sp và tora thermophila              

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    Polyporus sp và T villosa      20%, S communecho  [39]

- azinobis(3-ethylbenzothiazoline-6-  màu Reactive blue, Rbl5 và Ry70%, 42% và 21%.

[42] mangan perGanoderma lucidum KMK2 

AgaricomycetesBasidiomycota[36] Trong quá trình sinh

Sathiya và cs [ ] 45 th c hi n l  u tiên, t hi u qu kh màu trong nghiên     c u cao nh t 56.86% trong 5 ngày v  c th i ch a thu c nhu m ho t tính       t v i nghiên c u c a Shanmugam và cs[48]     ch ra r ng, có t     a thuc nhuc th i b kh b i laccase t     P florida sau 24h Mt nghiên c u  m i nh t c a Krishnaveni[35]    ng ch ra r ng, laccase t   P florida

 giá      môi ra   làm phân bón  có là

thu laccase  P floridalâu dài  giá thành 

Các loại enzyme được sử dụng:

   -acrylamid (Merck -   -azino-bis(3- ethylbenzthiazoline-6-sulphonic acid) Sigma Veratryl alcohol(3,4-Dimethoxybenzyl , alcohol) Sigma,NH4NO3, (NH4)2SO4 , MgSO4 , KCl, FeSO4, CaCO3,HCl, NaOH,

Các loại thuốc nhuộm được sử dụng:

Reactive blue 19 (Rb19); Reactive red 261 (Rr261); Reactive black 5 (Rbl

Reactive Yellow 186 (Ry186 Reactive blue 21 (Rb21) ); 

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Mho ng enzyme c n thi   trong 1 phút 30 0 C trong dung dm acetate 0.2M, pH 4.5

M u thớ nghi m g  m acetat natri 0.2M pH 4.5, b sung 100 àl phenol 

 0.1%, 20àl MnS04 5mM và 360 àl enzyme M u ki m ch ng g      m acetat natri 0.2M pH 4.5, b   0.1%, 20àl và MnS0 4 5mM h n h p này   30 0 C trong 10 phỳt B sung 20 àl H 2 O2 5mM Tru h n h p Xỏc  

nh ho mangan peroxydase b  h p th ánh sáng c a dung d ch    

M u thớ nghi m g     m tartrate natri 0.162M, pH 3, b sung 40 àl  veratryl alcoolol 60 mM và 400 àl enzyme h n h p này 30    0 C trong 10 phỳt B  sung 20 àl H2O2  b u ph n ng Tr  u h n h nh ho lignin peroxydase b  h p th ánh sáng c a dung d ch     c sóng 310 nm sau 1 phút

, 4, 5, 6, 7 và8 (trong citrate- phosphate0,1M và sodium phosphat 0,2M)

 2 và H2 màu trong máy Lovibond Checkit Direct COD vario thông 

n trong cái nút gasket trong chai

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Các

Ly tâm 200 0.000 vòng/phút, 4 0  gian 15 phút menem30 kDa hu T 

  0 C  Ly tâm  10.000 vòng/phút, 4 0   Thu 1,8 lít d ,

sang nuôi trong  veratryl alcohol  0 C

và do enzyme ng canh

Các ch ph m enzyme   bao gm:laccase tái t h p t  A niger

D15#26lcc1, h enzym t canh tr  ng Phomopsis sp.N7.2 trên môi tr ng basal,  laccase thi Mooyuan, h enzym chi t t bã giá th   P florida và P eryngii

 Enzyme c a bB sau nuôi  ã trng ca P florida và P eryngii: bãtr ng n m  c nghi n nh và   tách chit trong v i      thu nh n ch ph  m enzyme

Comment [C3]: Mình có phân tích các enzym khác không?

 Laccase Ch phm d ng b t l  accase A niger D15#26 1 lcc dng b t  và laccase

 c nghim   thu dung dch enzyme

4 A niger D15#26 1lcc 2.20 U/ml 0.00 U/ml 0.14 U/ml

Phomopsis sp N7.2 0.12 U/ml 0.00 U/ml 0.00 U/ml

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 khác nhau tính theo laccase: 0.05; 0.1; 0.2;

(A: Laccase P Eryngiieryngii; B: Laccase A Niger D15#26 1lcc ; C:

LaccaseP.florida ; D: Laccase ; E: LaccasePhomopsis sp N7.2)

          P Floridaflorida, P eryngii, Laccase laccase Mooyuan, P eryngii và  A niger D15#26 1 lcc

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Rr19 Rb21 Ry186 Rr261 Rb5

KHẢO SÁT KHẢ NĂNG KHỬ MÀU THUỐC NHUỘM CỦA ENZYME TỪ BÃ

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 citrate  photphat 0,1M pH 3, 4, 5, 6, 7 và phosphast

Natri 0,2 M pH 3, 4, 5, 6, 7 và pH 7, 8 0  ra trong Hình hình 3.4568

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Hình 3.6810.H các pH khác nhausau 24h.

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àu hoàn hoàn (pH5 92%; pH6 100%; pH7 100%) So

 (pH 4-6) (pH 5)  enzym có khpH  là

Nhi  có  ng l n kh n ng lo i m c    àu ác lo i thu c nhu m c a     enzyme t bã n m sò  P florida (Hình 3 7911) nhi  30 o C, hi u qu  kh màu 3 lo i  c ba lor i thuc nhu m  th nghi m  t giá tr l n nh t (Rb19 b kh màu hoàn      toàn; Rr261 b  kh màu 68% và Rbl5 b  kh màu 65%).Tuy v yK k hi nhi gim

60 xung t i 20 o C hoi 40 o C, enzyme v n có  kh  màu thu c nhu m   c a enzyme gi m, ch    t 19%, 37%, 82% ho     ng v i các  thuc nhu m Rbl5, Rr261 và Rb19 

 nhi 20 o C,40 o C, 50 o C hi u qu x lý 3 lo i màu thu c nhu      h n so v i  30 o C, ví d   u ki n nhi  20 o C, hi u qu x lý màu thu c nhu    m

Rb19 ch có 83%; Rr261 là 37%, Rbl5 là 19%   nhi 40 o C; 50 o C, hi u qu x   lý màu các lo i thu c nhu m gi m xu    ng m c dù enzym ho  ng t i pH này

u này có th do enzym không b n nhi     này, do v y ho t tính t ng th     gim.Kh màu t t 30 o C là m t y u t r   t thu n l i trong x  c th i b  thích v i nhi  ng  c ta.Tuy v y kh    màu t i các nhi  th(20 o C và 40 o Cc hi n các ph n ng kh màu v  

T các bình tam giác 100 ml 

trình bày trong hình 3.8 1012và 3.91113

1:  DO3.567 mg/l; 3: DO 6.86 5 mg/l; 4: DO 7.11 mg/l

DO 3.67 5 mg/l; 3: DO 6.86 5 mg/l; 4: DO 7.11mg/l

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A: laccase 0.1 U/ml; B: laccase0.2 U/ml; C: laccase 0.5 U/ml; D: laccase 1 U/ml)

Comment [C15]: Mình có phân tích các enzym khác không?

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Rb19 Tuy nhiên  chi phí enzyme khá cao         

( oxy hòa tan 3.56 mg/l; 2: là là 6.48 mg/l; 3: a tan 7.12 mg/l)

Hình 3.1 5.346 enzyme khác nhau sau 24h

ARb BRr them ghi chu

(Hình 3.1 ) 68 Các ,  màu kém Na2SO3 và Na2SO4

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 dòng vào vào Q = 225ml/h (9.72mg/h) ), tT

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NTRb  dòng vào là 225 ml/h

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 .2 QCVN 13 : 2008/BTNMT - Quy chu n k   thut qu c giav   c th i công nghi p d t may   

COD và BOD5 13 : 2008/BTNMT M màu (Pt-Co) cc thc gi m nhi u khi s d ng laccase t     P florida(t

2176 xun cao g p 2 l n cho phép v i tiêu chu n B     y vic kim soát s  kh màu thu c nhu   c th i m i ch     m b o thu c   nhu c kh màu Màu mang l i do các y  u t    c x lý Tuy  nhiênnghiên cchc kh   c th i d t nhu m nh     laccase và g i ý kh   t này v i h  thng x lý ti  hoàn thin h thng

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1 n và kh o sát kh    màu thu c nhu m c a 05 ch ph m enzyme      t  ng n m m c và bã th i tr ng n m Enzyme thu nh      c t bã thi P

florida có kh   màu nhi u lo i thu c nhu m: Rb19: 93%; Rr261: 66%;     Rbl5: 64%; Rb21: 48%; Ry186: 22%

2 u ki n ho ng ch y u c a ch ph m enzyme thô t       bã th i n m P florida Enzym hong ti nhi 70oC và pH 5 v cht ABTS, t i nhi  30oC và pH 5 7 v i thu c nhu m RB19    

3 Quá trình kh   c thi d t nhu m v   c th i NTRb trong h   thng x lý phòng thí nghi nh vu ki n sau: Q 

= 300 ml/h, n laccase: 0.20 U/ml, nhi 30oC, pH 7, DO = 7.3 mg/l, thi

  c th i trong h th ng 3.3 h H th    t hi u qu x lý BOD   

    màu Pt Co 85.8%, TSS 97%, t i tr ng 310 g thu c     nhu  c th u ra phù h p v i tiêu chu n B c a QCVN 13 :    2008/BTNMT

1 1 Ti p t c nghiên c u kh    c th i d t nhu m b   ng c a  n m P.floridavà tu ki n c a quá trình kh màu.  

2 2 Thu nh n h enzyme t giá th     trng nm P.florida và thu nh ng chng n m sò này qui mô l     s d ng cho h th ng x    c th i d t   nhum.

2 3 Tip t c nghiên c u kh      nh h enzyme laccase t giá th tr ng n m     

P.florida ho c c nh h s i c     a P.florida kh c th i d t nhu m nh   m t  d ng enzyme hay ch ng 

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1 Hoàng Th   tài c p S : Nghiên c u công ngh x      lý nguc th i gi t t y nhu   m  gi t-t y-nhu m t i Hà N i nh   m gim thi u ô nhi ng", S KH & CN - UBNDTP Hà N i, Hà N i   

2 ng Th C m Hà (2012), Nghiên c u công ngh s n xu enzyme ngo i bào      t  laccase, manganese peroxydase, lignin peroxydase (MnP, LiP) t vi sinh v t ph  c v x lý các ch t ô nhi      , Vin Khoa h c và Công ngh   Vit Nam, B Khoa hc và Công ngh 

3 Hunh Trung H i và Ph m Minh Huy n (1996), "Nghiên c u x       c thi

n Nhu m, H i th o khoa h c: Gi m thi u ch t th i trong công nghi p          d t", Hà N i.

4 H  Nghiên c u x lý m t s    cht m u h  c thi nhu m bt", Lu n án ti  n Hóa h c.

5 Cao H ng và Hoàng Th      ,

6 Nguy n H u Xuân Nhân (2010), Nghiên c u th c nghi m x lý màu ho t tính       

 c th i d t nhu m b ng mô hình công ngh sinh h c k khí hai giai       

7 ng Tr n Phong (2004),  ng trong d t nhu m  , Nhà xu t b n   khoa h c và k  thut Hà N i.

8 Nguy tài c p S : X lý các h p ch t h u có       khó phân h y trong  c th i d t nhu m b ng các quá trình ozon hóa nh m gi     m thiu ô nhi m t i Cty D t Minh Khai", Vi n Hóa h c -      Vin KH & CN VN, Hà

9 B Nông Nghi p và Phát Tri n Nông Thôn (2012), Th c tr ng và gi i pháp phát       trin s n xu t n m t i các t nh phía B c      

10 Lâ c th và công nghi p - Tính toán thi t k

11  t qu nghiên c u kh  o sát thuu tra c   

12 Nghiên c u th c nghi  xut công ngh kh thi x  c th i ngành công ngh d t-nhu m    , Lui Hc Bách Khoa Thành Ph H Chí Minh  

13 Abadulla E., Tzanov T.,Costa S., Robra K., Cavaco-Paulo A and Georg M Gübitz1 (2000), "Decolorization and Detoxification of Textile Dyes with a Laccase from Trametes hirsuta", Applied and Environmental Microbiology,

14 Archibald F S., Bourbonnais R., Jurasek L., Paice M G and Reid I D (1997),

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(2003), "Kraft pulp biobleaching and mediated oxidation of a nonphenolic substrate by laccase from Streptomyces cyaneus CECT 3335", Applied and Environmental Microbiology, 69: 1953-1958

16 Asgher M., Kamal S and Iqbal HMN (2012),"Improvement of catalytic efficiency, thermo-stability and Dye decolorization capability of pleurotus ostreatus IBL-02 laccase by hydrophobic Sol Gel entrapment", Chemistry Centre

17 Banci L., Ciofi-Baffoni S and Tien M (1999), "Lignin and Mn peroxidase- catalyzed oxidation of phenolic lignin oligomers", Biochemistry, 38: 3205-3210

18 Bortone G (1995), "Effects of an anaerobic zone in a textile wastewater treatment plant", Water Science and Technology, 32: 133 140 

19 Campos R., Kandelbauer A., Robra K H., Cavaco-Paulo A.and Gübitz G M

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20 Chandel A K., Kapoor R K., Singh A and Kuhad R C (2007), "DetoxiWcation of sugarcane bagasse hydrolysate improves ethanol production by Candida shehatae NCIM 3501", Bioresource Technology, 98: 1947-1950

21 Chivukula M and Renganathan V (1995), "Phenolic azo dye oxidation by laccase from Pyricularia oryzae", Appled Environmental Microbiol, 61: 4374

22 Chung K T and Stevens S E (1993), "Degradation of azo dyes by environmental microorganism and helminths", Environmental Toxicoloxy and Chemmistry, 12: 2121 2132 

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26 Gorman M J., Dittmer N T., Marshall J L and Kanost M R (2008),

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27 Guillen F., Gomez-Toribio V., Martinez M J and Martinez A T (2000),

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31 Jolivalt C., Raynal A., Caminade E., Kokel B., Le Goffic F.and Mougin C

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32 Junghanns C., Moeder M., Krauss G., Martin C and Schlosser D (2005),

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34 Kiiskinen L (2005), Characteration and heterologuos production of novel laccase from Melanocarpus albomyces, Helneski University of Technology,

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35 Krishnaveni M and Kowsalya R (2011), "Characterization and decolorization of dye and textile effluent by laccase from pleurotus florida- a white-rot fungi",

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38 Lee F (1997), "Reactive dyes and the problem if effluent",Journal for Asia on Textile & Apparel, (8(1)): pp 66-64

39 Lee I Y., Jung K H., Lee C H and Park Y H (1999), "Enhanced production of laccase in Trametes vesicolor by the addition of ethanol", Biotechnology Letters,

40 Linke D., Bouws H., Peters T., Nimtz M., Berger R G and Zorn H (2005),

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41 Nguyen Thi Phuong Mai, Le Quang Hoa, To Kim Anh, Duong Thi Phuc Hau and Dang Thi Thu (2011), "Optimization of the expression of laccase from white-rot fungus Trametes versicolorin Aspergillus niger D15#26", The 3rd AUN/SEED

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45 Sathiya Moorthi Perumal, Deecaraman Munuswamy, Periyar Selvam Sellamuthu, Murugesan Kandasamy and Kalaichelvan Puthupalayam Thangavelu (2007)

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