Tap chi Khoa hpc va Cdng nghe 49 (1) (2011) 111-118
ANH HU'6'NG CUA CH^
P H A MVI
K H U A NLACTIC LEN SW THAY D 6 | H A M Ll/aNG AXIT AMIN CUA CA TAP SAU THQI GIAN
BAO QUAN VA CPNG DUNG CHO NUOI TRONG THUY SAN
Nguyen The Trang, Tran Dinh Man, Pham Thanh Ha
I ien Cong nghe sinh hoc, I ien Khoa hpc vd Cong nghe Viet Nam Lien he vdi tdc gid: thetrang67((i>yahoo.com
Den Tda soan ngdy 25 thdng 10 nam 2009 1. DAT VAN DE
Vi khudn lactic tir xua da dugc diing nhieu trong bdo qudn thuc phdm, nhirng sdn phdm nhu axit lactic, axit acetic, H2O:, CO2 vd cdc bacterioein sinh ra bdi loai vi khudn ndy cd the lic chl nhiing vi sinh vat gdy benh, gdy hdng, keo ddi thdi gian bdo qudn vd cai thien sy an todn cua sdn phdm [4].
Cd Id ngudn cung cdp protein rdt quan ttpng cho con ngudi, tuy nhien cd cd ban che Id dl uon, hdng hon cdc loai thyc phdm khdc vi nd chiia mdt lugng ldn axit amin ty do. Cd thudng dugc bdo qudn bdng che phdm vi khuan lactic khi viec ldm khd cd gap khd khan do thdi tiet am udt hoac viec ldm ddng lanh hay che bien Id qud tdn kem [2]. Trong thiic an chan nudi gia siic vd nuoi ttdng thuy sdn cd Id thdnh phdn quan trgng khdng the thilu. Bdt cd Id ngudn protein chdt lugng cao, dl tieu hda. Thuc dn cho chdn nudi gia siic vd nudi thuy sdn d nude ta can khodng 300.000 tan/ndm, ttong dd bdt cd sdn xudt cdng nghiep chi ddp ling 1/10 nhu cdu, do vay phdi nhap khdu [11]. Nghien ciiu sdn xudt thuc an cd chdt lugng cao theo quy md nho nham tan dung nguyen lieu cd tap phdn tdn, tang khd nang ddp ling nhu cdu bdt cd cdng nghiep, gidm nhap khdu la cdn thiet [6].
2. VAT LIEU VA PHlTONG PHAP 2.L Vat lieu nghien cmi
Chung vi khudn lactic: Pediococcus pentosaceus HN02; Lactobacillus brevis HN26;
Lactococcus lactis subsp. lactis HNU vd Lactobacillus delbrueckn subsp. delbrueckii HN34 ttong bd suu tap chiing cua Vien Cdng nghe sinh hpc. Cdc chiing vi khudn lactic dugc nhdn gilng rieng re tren mdi trudng MRS [3], d nhiet dp vd thdi gian thich hgp. Djch nudi cua bing chung dugc ly tam thu sinh khdi vd trdn vdi chdt mang. Cdc chl phdm vi khudn lactic bdo qudn ca dugc xdc dinh LD50 tai Vien Kilm nghiem Bd Y tl cho kit qud khdng sinh ddc td.
Cd tap nude man duae mua d cha cd Dd San (Hdi Phdng) dugc thilt kl bdo qudn cd theo 4 16 thi nghiem (TN) sau: mdi Id bd sung 2% chl phdm, 2% glucoza, 0,5% NaCl, 0,1% axit sorbic.
Cdc Id TN bd sung cdc dang chl phdm nhu sau: TN 1: Chi cd chat mang, khdng bd sung vi khudn lactic- TN 2: Chl phdm chua hdn hgp 2 chiing HN02 vd HN26; TN 3: Che phdm chiia
Nguyin Thi Trang, Trin Dinh Min, Ph^m Thanh HS
hon hgp 2 chiing HN11 va HN34; TN 4: Chl pham chira hon hgp 4 chiing vi khudn lactic tren.
Sir dyng chat phy gia Soya Lecithin lam chat keo dinh cho thue dn nudi trdng thuy sdn.
2.2. Phuong phdp nghien ciru
Cac mau ca dugc thuy phan bdng HCI 6N chua 0,5%) phenol, ii d I IO"C trong 24 gid. Dung djch thiiy phdn dugc Idm sach bang each ly tam Ipc bd cdn. Dung djch mdu dugc ldm khd chdn khdng. Can khd dugc hda tan trong HCI 0,IN. Phdn tich axit amin tdng sd vd axit amin ty do bdng sdc ki tren may phan tich axit amin ty dpng HP-Amino Quan Seriese ll cua Vien Cdng nghe sinh hpc [7]. Ddnh gia chat lugng nude ao nudi vd chat lugng cd theo TCVN [9, 10].
3. KET QUA VA THAO LUAN
3.L Anh hudng ciia che pham vi khuan lactic den hdm lugng axit amin ciia ca bao quan BSng 1. HSm lugng axit amin tdng s6 trong c^c m§u cS dugc b^o quin sau 25 ngiy
STT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
Axil amin Aspartic axit Glutamic axit
Serine Histidine
Glycine Threonine
Alanine Arginine Tyrosine Cystein+Cystine
Valine Methionine Phenylalanine
Isoleucine Leucine
Lysine Proline Tdng sd
Ham lugng axit amin tdng sc") ("o (g axit amin/100 g mau)) Ddi chung
0,32 0,69 0,12 0,01 0,45 0,09 0,32 0,27 0,10 0,07 0,17 0,09 0,14 0,13 0,28 0,54 0,11 3,90
TN 1 0,15 0,38 0,06 0,01 0,34 0,07 0,31 0,17 0,16 0,04 0,15 0,06 0,07 0,11 0,20 0,36 0,10 2,74
TN2 0,23 0,51 0,09 0,01 0,38 0,08 0.33 0,20 0,18 0,05 0,17 0.07 0,09 0,13 0,21 0,39 0,11 3,23
TN3 0,22 0,51 0,09 0,01
TN4 0,24 0,55 0,10 0,01
0,39 0.42 0,08
0.33
0,08 0,33
0,20 0,22 0,18
0,05 0,16 0,08 0,11 0,12 0,22 0,45 0,10 3,30
0,16 0,06 0,17 0,08 0,11 0,13 0,22 0,45 0,10 3,43
Day la edch bdo qudn cd bang phuong phdp len men vi sinh vat. Ca tap ttdn vdi che pham cho vdo cdc thiing nhya ii kin. Trong qud trinh u chua, axit lactic dugc tao ra lam pH gidm, sdn
Anh hw&ng cua chi phSm vi khuin lactic ISn sw thay ddi hSm Iwang axit amin...
phdm dugc bdo qudn hang thdng khdng bj hdng. Cd ddng l^nh Idm doi chirng vd cd thi nghiem sau 25 ngdy dugc xdc djnh axit amin tong so. Kit qud dugc trinh bdy d bdng Ivd sdc ki do d hlnhl.
DM>1 • •••-SR.MnB*-3M«(CT«».l.l»
[LJjkJiilk^iMJL
I;'rai: Z3JZ
1
t E k*.
IAJAAAM^
a. Cd bao qudn dong lanh b. Cd bao quan bSng che phdm khong chOra vi khuan lactic
jJLMl-ijfciii^^
.:_iJ
c.Ca bao quan bdng chl phdm chiia hin hop 4 chung vi khuan lactic Hinh 1. Sic ki do axit amin tong so
Kit qud d bdng 1 cho thay so vdi mdu cd bdo qudn bdng ddng lanh hdm lugng axit amin ting sd trong cdc mdu thi nghiem diing chl phdm vi khuan lactic cd gidm (axit amin tdng so 3,23 3,43 (g axit amin/100 g mdu) so vdi 3,9 (g axit amin/100 g mdu)), tuy nhien vdn ddm bdo yeu cau dinh dudng cua sdn phdm [9]. So sdnh cdc thi nghiem thdy cdc mdu cd bdo qudn bang chl pham vi khudn lactic diu chiia hdm lugng axit amin tong so cao ban mdu ed bdoqudn bang chl phdm khdng cd vi khudn lactic. Protein trong cd bj phdn buy chii yeu do enzim sdn ed trong rupt cd vd do vi sinh vat xdm nhap vdo cd gay ra. Khi bo sung vi khudn lactic vdo cd, axit lactic sinh ra nhilu ldm gidm pH cua sdn phdm, ban chl sy boat ddng cua enzim phdn buy vd sy phdt triln ciia vi sinh vat gay thdi. Nhimg bacterioein sinh ra bdi vi khuan lactic cung gdp phdn ire chl nhimg vi sinh vat gay benh vd gay hdng, ldm cho sdn phdm an todn yd giir dugc lau ban [8].
Mdu cd bdo qudn bdng chl phdm chiira cd 4 loai vi khudn lactic Id tot nhdt. Hdm lugng axit amin con giir lai trong sdn phdm Id cao han 2 thi nghiem chi bdo qudn bang 2 loai vi khudn lactic. Khi kit hgp sir dung cd 4 loai vi khudn trong bdo qudn, hdm lugng axit sinh ra nhieu ban va nhung chdt khdng khudn cQng da dang ban do dd cd khd ndng bdo qudn cd tdt han. Ket qud ndy phii hgp nhimg nghien cuu trudc day [8].
Kit qua phdn tich axit amin ty do dugc ttinh bdy d bang 2 vd sdc ki dd axit amin ty do cua mlu ddi chirng, TN 1, TN 4 d hinh 2.
Tir bdng 2 cho thdy cdc mdu cd dugc phan tich cd gdn dhy dii cdc axit amin (chi co 4-Hydroxy Proline khdng thdy tdn tai trong cdc mdu cd). Vi cd chiia mdt lupng Idn axit amin ty do nen nd dl hu hdng ban cdc loai thyc phdm khdc. Trong dd cd 8 axit amm thiet yeu la isoleusine, leucine, lysine, methionine, phenylalanine, threonine, fryptophanva valine ca the dong vat khdng tu tdng hap duae md bdt bupc phai dugc cung cap tu thyc pham ben ngoai [2].
Nguyin Thi Trang, Trin Dinh Min, Pham Thanh Ha
A.\it amin histidine c6 o ca ddng lanh nhung khdng thay cd a nhirng mdu cd thi nghiem. Trong khi 2 axit amin khac la asparaginc va glulamine lai thay co mpt lugng nhd d cdc mdu cd thi nghiem nhung khdng tim ihav trong ca tmri ddng l^nh, ham lugng mpt sd axit amin tu do d cdc mail thi nghiem tham chi con lang len so vdi man ca luai ddng lanh. Vi vay, axit amin tu do tdng so o cac mau thi nghiem cao hirn o mau ca luai ddng lanh. Ddc bipt o thi nghiem cd bdo quan bang clic pliain chira ca 4 loai vi khiuin lactic hiiin lugng axit amin ty do tdng sd len tdi 61,4 % (mg axit amin/100 g mau) Irong khi man ca luai chi c6 52,()3"(. (mg axit amin/100 g mau), cao ban gan 1,2 lan. I)a\ Id uu diem ciia phuang phdp bao qudn cd bang che phdm vi khudn lactic.
Bani; 2. Ham lugng a.xit amin ty do trong cac man ca dupe bao quan sau 25 ngay
STT 1
-»
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
.\\\{ amin
Aspartic axit Glutamic axit Asparagine Serine Glutamine Histidine Glycine Threonine Alanine Arginine Tyrosine
Cystein+Cystine Valine
Methionine Tryptophan Phenylalanine Isoleucine Leucine Lysine 4 - Hydroxy Proline Proline Tdng sd
Ham lugng axit amin tu do (% (mg axit amin/100 g mdu)) Ddi ehimg
0.72 3.62 1,33 0,45 5,20 1,02 4,15 17,06
3,02 0,27 2,56 1,17 1,63 1,41 1,48 3,29 3,47
0,18 52,03
TN 1 0,04 0,58 0,04 0,31 0,18 1,54 0,35 8,06 15,14 10,08 0,41 3,10 1,71 0,85 2,78 0,91 3,10 3,34
0,40 52,92
TN2 TN3
0,10 ^ 0,05 0.74
0,01 0,20 0,10 0,82 0,33 8,24 16,20 12,80 0,49 3,64 1,83 1,14 3,55 1,13 3,69 2,64
0,46 58,11
0,60 0,06 0,26 0,21 1,46 0,37 8,08 14,15 11,06 0.39
TN4 1,04 0,98 0,09 2,28 0,26 2,36 0,64 8,09 16,00 12,16 0,52
3,12 3,36 1,75 '. 1,78
0,90 1,26
2,86 ; 2,63 0,94
3,12 3,37
0,50 53,25
1,68 3,36 2,39
0,52 61,40
Anh hw&ng cua chi phim vi khuin lactic ISn si/ thay ddi hSm Iwpng axit amin.
Ca bao quan dong lanh b. Ca bao quan bang che phSm khong chua vi khuAn lactic
c.Ca bao quan bang che pham chira h6n hap 4 chiing vi khudn lactic Hinh 2. Sac ki do axit amin ty do
Dimg L. plantarum vd L bulgaricus Idm gidng khdi ddng len men sdn phdm cd len men. 3 giong vi khudn lactic (L. plantarum, P pentosaceus, L. mesenteroides) tdch tir sdn phdm cd dugc dua vdo thir khd nang bdo qudn cd trong vdng 52 ngdy d 8°C [4; 5]. Ket qud cho thdy trong trudng hgp L. mesenteroides dugc lya chpn cho ket qud tot nhdt: nhirng thay ddi trong thdnh phdn hiiu ca dat yeu cdu, hdm lugng histamine sinh ra thdp nhdt, sdn phdm co khd ndng bdo quan trong thdi gian ddi. Cd bdo qudn ddm bdo ve hdm lupng axit amin, ddc biet Id nhu-ng axit amin khdng thay the nen da dugc sir dung Idm thirc dn nudi trdng thuy sdn giup ngu ddn chu dpng gidi quylt dugc nguon thOrc dn du thira sau ddnh bdt ciing nhu khan hiem vdo miia bien dpng [2].
3.2. Anh hudng ciia chl phim vi khudn lactic den cam quan ciia ca bao quan
Ddnh gid cdm quan chat lugng cd [9] cho thay: Ci mau bdo qudn ddng lanh: Cd rat tuai, vj tanh nhe. 6 TN 1: Cd dau hieu uan hdng, miii khd chju. 6 TN 2, 3 vd 4: Thjt cd khdng ed miii la, cau triic ca thjt vdn cdn tot. Diing chl phdm vi khudn lactic cd thl giiip bdo qudn ed tuoi sing md khdng Idm thay ddi miii vj sdn phdm. Ngodi ra cdc gdc ty do vd cdc chdt oxy hoa it xuat hien ban. Vi khudn lactic ngdy nay da dugc sir dung nhilu trong bdo qudn cdc sdn pham cd.
Ngodi nhiem vu bdo qudn dugc sdn phdm lau ddi chiing cdn cd khd ndng ban chl toi da cdc vi sinh vat gay benh nhu B. subtilis, B. mesentericus, Pseudomonas, P vulgaris, C. putrificum, C.
sporogenes. Salmonella, Sarcina, Streptococcus, Micrococcus, E. coli, ... vdi thdi gian bdo qudn dai md chdt lugng tuang ddi dn djnh, ddc biet axit amin trong cd hdu nhu thay dii khdng ddng kl [1; 8]. Ddy chinh Id yeu to quan trpng dl chl phdm lactic diing bdo qudn cd khdng thua gi so vdi cdc edch bdo qudn khdc nhu sir dung lugng dd rat ldn (thudng ti le 1 : 1) md lai kinh te ban.
3.3. iTng dung ca bao quan bdi che pham vi khuan lactic cho nuoi trong thuy san
Nguyin Thi Trang, Trin Dinh Min, Ph^m Thanh HS
Ca biln tap Ibutrng chi cd the su dyng trong thdi gian ngdn tir I - 2 ngdy. Viec sii dyng vi khuan lactic dl bdo quan giiip ngudi nudi dy trti dugc thirc dn cho cd. Trong nghien ciiu ndy cd sau 25 ngay bao quan bang die pham vi khuan lactic dugc diing tryc tiep ldm thiirc dn nudi cd Loc Bdng vd so sdnh vdi thue an bdng ca thdng thudng. Su dyng cd bdo qudn bdng vi khudn lactic lam thue an nudi trdng thiiy san dugc trinh bdy d hinh 3.
( a biu) qudn bang che pham vi khuan lactic sau 25 neav
C a dupe nghien vdi phu gia Soya Lecithin lam thirc Sn cho nuoi cit Hinh 3. Anh Cci trudc, sau b^o qu^n vd ldm thu-c Sn cho nudi ca
Ddnh gid dnh hudng ciia thirc dn lam tir cd bdo qudn bdng che phdm vi khudn lactic den mpt sd chi tieu cdm quan nude ao nudi trdng cd Lde Bdng nudi trdng thdm canh d Hdi Hdu, Nam Djnh trinh bdy d bdng 3 vd chdt lugng cd d bdng 4.
Bang 3. Anh hudng ciia thdc an tdi mpt so chi tieu cdm quan chit lu-gng ao nuoi
STT 1 2 3
Chi tieu Mdu eua nude ao
Dp trong cua nude
Miii ciia nude ao
Cd nghien cd vi khudn lactic Mdu xanh sdng hay xanh nhat cho biet nude ao cd mat dp tdo thich hgp, cd ddy dii dxy.
Anh sdng mdt trdi xuyen qua 30- 40 cm
Miii ddc trung ciia ao nudi
Cd nghien khdng cd vi khudn lactic
Mdu xanh thudng hay xanh hai tdi cho biet nude ao cd mat dp tao tuong ddi, cd dxy.
Anh sdng mdt ttdi xuyen qua 20- 30 em
Miii tanh cua tao phdt trien BSng 4. Odnh gid chclt lu-gng cd
STT 1 2 3 4 5
Chi tieu Mdu sdc Miii, vi Trang thdi Trgng lugng cd trung binh (g/con)
Nang sudt (%>)
Cd ao thi nghiem Mdu sdc sang ty nhien Mui tanh tu nhien cua ca,
khdng cd miii la
Ca thjt ttdng, chdc, dan hdi tdt 850 ± 5
113,3
Cd ao ddi chung Mau sac sang ty nhien Miii tanh ty nhien cua cd, khdng
cd mui la
Ca thit ttdn^ thudng, hai chdc, dan hdi trung binh
750 ± 5 100
Anh hw&ng cua chi phim vi khuan lactic len sw thay ddi hSm Iwang axit amin...
Tir bdng 3 vd 4 cho thdy diing ea nghiln cd vi khudn lactic lam thiic an cho ca khong lam 6 nhilm nude ao nudi, tham chi nd cdn cd tac dung iic chl sy phat triln eiia tao lam nude trong hon, dung ca nghiln cd vi khudn lactic lam thirc an co the giiip ca Idn nhanh ban, thjt cd chdc han, ndng sudt cd tang 13,3"o.
4. KET LUAN
Sir dung che phdm lactic diing bdo qudn ea cho kit qua tuang tu so vdi each bdo quan ddng lanh, diing che pham \ i khudn lactic co thl giup bao qudn ca trong thdi gian 25 ngdy van dam bdo chdt lupng, dac biet ham lugng axil amin it thay doi. Mau ea bdo quan bdng chl phdm chira cd 4 chiing \i khudn lactic P pentosaceus HN02; L. brevis HN26; L. lactis subsp. lactis HNI1 va L. delbrueckii subsp. delbrueckii HN34 la tdt han nhirng chl phdm co 2 loai vi khudn lactic.
Ci thi nghiem ed bdo qudn bang che pham ehua 4 loai vi khudn lactic ham lugng axit amin ty do tong so cao hon mdu cd tuai ddng lanh 1,2 ldn. Chl pham vi khudn lactic co thl giup bdo quan cd tuai sdng md khdng Idm iha> ddi miii \ i sdn phdm. Thirc an co vi khudn lactic bo sung 'do ao nuoi khdng Idm d nhiem nude. Cd nudi su dung thuc an tir san phdm cd bdo qudn bdng v' khudn lactic ldn nhanh. thjt cd chdc, ndng sudt tdngl3,3''o.
TAI LIEU THAM KHAO
1. Ahamed J., Mahendrakar N. S. Chemical and microbial changes in fish viscera during fermentation ensiling at different temperatures, Bioresource technology ISSN 0960-8524, 59(1) (1997) 45-46.
2. Bustari Hasan Fermentation of Fish Silage using Lactobacillus pentosus, Jumal Natur Indonesia ISSN 1410-9379 6 (1) (2003) 11-15.
3. De Man J. C , Rogosa M., Sharpe M. E. A medium for the cultivation of lactobaeili, J.
Appl. Bact. 23(1960) 130-135.
4. Larisa Glatman, Vladimir Drabkin, and Alexander Gelman Using lactic acid bacteria for developing novel fish food products, J. Sci. Food. Agrie. 80 (2000) 375-380.
5. L. J. Yin, C. L. Pan, S. T. Jiang - New Technology for Producing Paste-like Fish Products using Lactic Acid Bacteria Fermentation, Joumal of Food Science 67 (8) (2006) 3114-3118.
6. Nguyin Thl Trang, Trdn Dinh Mdn Sir dung vi khudn lactic Pediococcus pentosaceus HN02 dl sdn xudt chl phdm bdo qudn cd. Tap chi Khoa hpc vd Cdng nghe 45 (4) (2007) 35-42.
7. Phan Van Chi, Nguyen Bich Nhi, Nguyin Thj Ty Xdc djnh thdnh phdn axit amin bang phuang phdp ddn xudt hda vdi 0-phthadialdehyt (OPA) vd 9-fluorenylmetylchrolformal (FMOC) tren he HP-Amino Quant Series II, Ky ylu Vien Cdng nghe sinh hpc 1997, Nhd xuat bdn Khoa hpc vd Ky thudt Hd Npi, 1997, tt. 454-461.
8. Sanni A. I., Asiedu M., Ayemor G.S. Microflora and Chemical Composition of Momoni, a Ghanaian Fermented Fish Condiment, Joumal of Food Composition and Analysis 15 (5) (2002) 577-583.
9. TCVN 5090-2008 (2008). ISO 4121: 2003 Phdn tich cdm quan - Hudng ddn sir dung cdc thang do dinh lugng dac trung, 9 trang.
10. TCVN 1644-2001 (2001) Thirc an chdn nudi. Bdt cd, Yeu cdu ky thudt, 5 trang.
Nguyin Thi Trang, Trin Dinh Min, Ph^m Thanh HS
11. Cong nghe san xiial bpl ca chan nudi quy nio nhd tir hdn hgp nhieu loai cd tap, Vien Nghien ciru nudi lioiig thiiy san II.
http://\vww.vielli n 11. c o m.vn/ k i t hual/chiiyende/iesearch/sxholca.htm.
SUMMARY
THE CHANGIOl AMINO ACID AFTER FERMENTATION OF FISH SILAGE USING LiCTlC.lClD BACTERIA COMPOST AND APPLICATION IN AQUACULTURE
Lactic acid femientation is ulili/cd throughout the world as a prime method of preserving fresh food.
Fish, however, is more perishable than other meats as it contains relatively large quantities of free amino acids. If llsh is not preserved, il will quickly spoil. The lactic acid bacteria (LAB) are added lo raw fish due to the formation of lactic acid. In these products, the pH of fish mixture is lowered making the product .safer and easier to keep. The suitability of LAB for fish silage fermentation was nucstigatcd. Four kinds of compost were the respectively prepared, using without LAB; w itli two of LAB strains; with the other two of LAB strains and with four of LAB strains as fermentation starter. The starter were added to raw fish and stored at room temperature for 25 days. Changes in amino acids of silage after storage were evaluated, and the values were compared to the frozen fish. There were no specific changes in the level of amino acid within 25 days of the storage. Total amino acids for frozen fish and for fermented silages were added with LAB strains were 3.90 and 3.23"b 3.43"o (g amino acid 100 g sample), respectively. Total free amino acids were 52.03 and 58.11% 61.40%) (g amino acid/ 100 g sample), respectively. Fermented fish silage can be successfully prepared using fermentation starters of complex of four of LAB strains P pentosaceus HN02; L. brevis HN26; L. lactis subsp. lactis HNI 1 and L. delbrueckii subsp. delbrueckii HN34 w ithin 25 da\s of storage.
Keywords. Lactic acid bacteria, amino acids, compost, fish, preservation.