Tran VSn Chi va Dtg Tap chi KHOA HOC & CONG NGHE
NGHIEN C i r u K H A NANG LEN MEN CUA T O HgfP NAM MEN VA VI KHUAN L A C T I C TlT NGUON THAN CAY NGO, R O M T I / O I VA C O L A M T H i r c AN GIA SUC
Tran Van Chi', Pham Thj Tuylt Mai, Nguyin Xuan Ninh, Dinh Thi Kim Hoa, NguylnThj Huong Trudng Dai hoc A dng Ldm - DH Thdi Nguyen TOM TAT
Sir dung to hpp chiing nam men va vi khuan sinh lactic d€ ISn men than cay ngo, ram tirai va co
\hm thirc an gia siic nham tang s6 lupng vi sinh co loi cho du6ng ruot, h6 trg he tieu hoa ciia dpng vat an CO duoc t6t ban. Viec len men dupc thuc hien theo 2 pha - ban ran va long, qua do sinh khoi nam men sau pha hieu khi tang den lO', nhung sau pha y^m khi sinh kh6i giam con 10^, Sinh kh6i cua vi khuin lactic tSng din trong 2 pha len men, kat thuc pha 2 s6 lupng ti b^o vi khuin dat khoang 10'° Qui trinh len men c6n i^ su chuydn hoa sinh hpc nguyen lieu thuc vat thanh protein don bao, lam tang ham luong protein len 53,1 %. Hon nCra vdi sir chuy§n hoa sinh hpc nay da lam thay doi th^nh phan acid amin trong nguyen lieu, t§ng ham lupng acid amin khong thay thi Tir khoa: ndm men, vl khudn lactic, len men
DAT V A N D E
Viec tang mat dp nuoi gia sue lam gia tang b?nh tat do bj nhilm cac mam benh khac nhau, dac biet la vi khuan duang ruot. Chiing lam giam nang suit va la nguon nhiem tiem timg cho cac san phSm thit, giam an toan thuc pham eho con nguai. Bo sung probiotic trong thirc an dong vat da dugc nghien ciiu tir rat lau, vai muc tieu lam tang so lugfng vi sinh v^t CO lgi trong duong rugt. Trong nhieu th^p ky qua, khang sinh duge bo sung trong thire an de phong benh va kich thich sinh truong ciia v§t nuoi. Tuy nhien viec su dung khang sinh khong dung each da va dang gay ra nhilu moi lo ngai ve sy khang thuoc cho vat nuoi, ehinh vi vay viec cam sOr dung khang sinh ho§e giam ham lirgng khang sinh trong thirc an chan nuoi da dugc ap dung 6 nhieu quoc gia tren the giai [5].
Bien doi khi hau va 6 nhiem moi truong la van dl c4p thiet hien nay co anb huang den xu huang nghien ciru va phat trien khoa bgc cong nghe ciia toan the giai. Vdi xu huong phat trien ben virng, cac phirong phap cu gay 6 nhilm moi trucmg dan dan dugc thay the bing cac phuang phap mai than thien hon voi moi trucmg. Cac quy trinh cong nghe sach Tel-0965 051219, Email IranchiinfSgmail com
khong tao ra chat thai rat dugc quan tam boi cac nha khoa hoc, nghiC-n ciiu, quan ly va nha dau tu [3].
Dual sir huong dan cCia giao su, tien sT khoa hoc Borisenko E.G. da c6 04 luan van tien sT bao ve thanh cong trong 02 nam gan day, lien quan den viec sir dung nguyen lieu thue vat de san xuat thyc pham va thiic an gia sue giau dinh duang [1, 2, 6] - trong d6 eo xii ly ram ra va than eay ngo. Cong nghe sir dung a day la cong nghe sach vi tan dung nguon phu pham nong nghiep va khong thai ra phe pham nao trong qua trinh san xuat vdi nguon vi sinh vat duac phan lap tir ty nbien.
Cac nghien ciru trong nude hien nay bang phucmg phap cong nghe sinh hpc tap trung chu yeu vao tien xir ly ram ra va tao ra cac che pham cellulase co hoat tinh cao dl thuy phan ram ra va ien men thanh con nhien lieu [7]. Con than eay ngo dirac 0 vdi mat ri va ure dimg lam thiic an gia sue vdi gia tri dinh duong thap [2],
Mue tieu cua chiing toi trong nghien ciru nay la" Ung dung cong nghe len men vi sinh de ehuyen hoa mgt phan co chat tii' than cay ng5 thanh protein dan bao va cac chat cd Igi khac, dong thdi hinh thanh to hgp vi sinh vat trong san pham (probiotic).
HI
Iran Van Chi va Dig Tap chi KHOA HOC & CONG NGHE VAT LIEU, PHUONG P H A P NGHIEN CUU
Vat lieu nghien eihi
Than cay ngo, ram tiroi va eo dugc thu tir nhimg canh ddng tren dia ban tinh Thai Nguyen.
Cac ehung nam men (Pichia anomala 9a) dugc phan lap tir nguon ty nhien dugc km giiJ tai phdng thi nghiem Cong nghe vi sinh - Trudng Dai hge Ndng lam Thai Nguyen Chimg VI khuan sinh lactic: L.acidiphtUus La5, dugc phan lap tir sira ddng vat va dugc luu giu' tai phdng thi nghiem Cong nghe vi sinh - Trudng Dai hpe Ndng lam Thai Nguyen.
Phuang phap nghien cuu Phutmg phap xtr ly nguyen lieu
Than cay ngd, rom tuai va cd dugc thu tren dja ban tinh Thai Nguyen dugc nghien vdi vdi kich thirdc nbd (3-5 cm x 7-9 cm) dugc thanb triing va de ngudi trude khi nudi cay.
Phuong phap len men vi sinh v|t theo 02 pha:
Nguyen Heu sau khi dugc xir ly se dugc cay cac chiing vi sinh vat nghien cii'u (nam men va vi khuan sinh lactic) vdi sd lugng chung khdi dpng nhu nhau. Pha 1 thuc hien len men tren mdi trudng ban ran vdi do am 50 - 60%
theo doi su tang sinh ciia te bao nam men, te bao vi khuan trong 72 gid. Pha 2 chuyen tii' trang thai ran sang trang thai long bang each cho them nude (vdi ty le 3 nudc/1 khdi lugng nguyen lieu tirpha 1) de thyc hien len men yem khi ddng thai theo ddi sy thay ddi sd lugng te bao vi sinh vat va pH.
Phuang phap dem sd lugng tl bao vi sinh vat - Phuong phap pha loang va dem khuan lae:
lay I gram mau thf nghiem sau len men dem pha loang vdi cac cap do khac nhau (tii' 10"' - lO'") rdi cay len 02 loai mdi trudng dinh dudng khac nhau la Hansen va MRS de nuoi trong tii am, sau 48 gid dim sd lugng khuan lac mgc len tir dTa mdi tnrdng va xir ly ket qua.
Bang 1. Sii- tdng sinh cua le
Xac djnb bam lugng protein bang phuang phap kjeldahl.
Dmh lugng acid amine bang phuang phap phan tich: H/QT/I9.46 tren thilt bj HPLC.
KET QUA VA THAO LUAN
Anh hirdng ciia nguyen lieu den sur phat trien ciia vi sinh vat trong pha 1 Ddi vdi thiic an gia sue li ehua, ngudi ta thudng sir dung cac chCing len men lactic. Tuy nhien nlu chi sCr dung cac chiing vi khuin lactic thi dan thuan ehi giai quylt dugc van de thay ddi miii vi cua san pham cudi ciing.
Viec dua nam men vao nudi eay cimg cac chiing vi khuan lactic trong thi nghiem ciia ehiing tdi de giai quyet bai muc dich: 1 la khao sat sy ket hgp eiia chiing nam men nghien euu vdi chung vi khuan lactic, 2 la tan dung triet de ngudn dinh duotig trong co chit dl chuyen hda protein thuc vat thanh protein vi sinh vat (protein don bao).
Trong thi nghiem nay chiing tdi lya chpn chung nam men vdi muc dicb nhan nhanh de tang sinb khdi d pha dau tien (pha hieu khi- nudi tren mdi trudng ban rfin). Kit qua trong bang 1 cbo thay trong pha dau tien sd lugng te bao nam men tang len nhanh ehdng trong hai ngay dau tien (len den lO' te bao/gram), sang ngay thir 3 tde do tang cd phan ch&ng lai va cd chieu hirdng giam Dieu nay hoan toan tuan theo quy luat dudng cong sinh trudng cua vi sinh vat, tiic la bat dau sang ngay thii 3 sd lugng te bao nam men chuyen dan sang pha suy vong
Mac dil vi khuan lactic phat trien tot d dieu kien yim khi, tuy nhien theo bang 1 eo the thay ring, ngay d dilu kien hiiu khi vi khuin lactic van tang sinh va tdc do tang cham hon nhieu so vdi nim men. Ket thuc pha thir nhit lugng te bao vi khuan cung dat 10^ te bao/gram eanh trudng.
hdo vi smh vdt trong nuoi cdy pha 1
So tupng te bao nam men sau So Iirpng te bao vi khuan sau ' khoang thoi gian nuoi cay pha l(x khoang th6i gian nuoi cdy pha l(x
109/8 ram) IQ9/gram) Loai nguyen
liSu
24h 48h 72h 24h
1 Co 2 Rom tuai 3 Cay ngo
72h 3,1
2.2 4,5 3,8
4,3 3,7 5,9
0,3 0,2 0,3
0,4 0,3 0,5
0,6 0,5 0,7 112
Tran Van Chi vd Dig T9p chi KHOA HOC & CONG NGHE 134(04): 111-114 Anh huffng cua nguyen lieu den su* phat trien cua vi sinh vat trong pha 2
Vi?c tien hanb thi nghiem trong pha 2 dugc thyc hien trude tien la chuyin tir mdi trudng ban ran sang mdi trudng ldng bang cdch bd sung mgt lugng nude nhSt dinh, lam sao de nude ngap len tren be mat mdi trudng, vda de pha loang lugng te bao vi sinh vat ddng tbdi vira thiet lap trang thai len men yIm kbi.
Sd lieu bang 2 cho tbay: sd lupng te bao nam men tiep tue giam, sau 72h theo doi luang te bao nam men giam khoang 100 Ian (tii' lO' xudng lO'te bao/ml). Tuy nhien sd lirgng te bao vl khuan lactic lai tang len nhanh chdng. Theo doi 72 gid nudi trong pha 2 (pha ldng) thiy rang lugng tl bao vi khuan tang len din lO'** tl bao/ml. Va cd the thiy rang b4t dau sang ngay thii' 3 su tang sinh khdi ciia vi khuin cham lai va cd xu budng giam. Cd nghTa la sau 72h len men yem khi cd the ket thiic qua trinh de hoan thien san pham.
Bang 2. Sir Idng sinh ciia ti bdo vi sinh vgt trong nuoi cdy pha 2 TT
1 2 3
Lo^i nguyen li^u
Co Rom mai Cay ngo
So lirong te b^o nam khoang thoi gian nuoi
l(x lO'/ml) 24h
0,7 0,6 1,2
4Sh 0,2 0,1 0,5
men sau i cay pha
72h 0,07 0,03 0,09
So iuo'i Ithoang
24h 1,5 1,4 1,9
[Ig t^ b^o vi lihuSn sau thd'i gian nudi cay pha
l(x lO'/ml)
48h 72h 7,3 7,2 7,2 7,0 10,2 9,3 Su thay doi pH ciia canh trudng trong qua
trinh nudi cay pha 2.
De danh gia dugc bieu qua len men cua vi khuan sinh acid lactic, ehiing tdi tien hanh xae djnh pH ciia cac mau thi nghi6m theo mdt khoang thdi gian nhat dinh.
BSng 3. Sir thay doi pH cua moi tneang khi nuoi cdy trong pha 2
Bang 4, Sir thay doi hdm luong protein vd acid gmln trong cdc mdu thi nghiim
TT 1 2 3
Loai nguy€n
li€u Co Rom tuoi Cay ngo
S(i- thay a6i pH theo thdi gian 24h 5,3 5,0 5,1
48h 72h 4,2 3,5 4,1 3,4 3,9 3,3 Ket qua bang 3 eho thay: pH ciia cac mau thi nghipm theo thdi gian giam din. Dilu nay khang dinh lai mgt lan nii'a kit qua ciia bang 2 la hoan toan dimg. Trong dieu kien yem khi vi khuan lactic vCra tang sinh khdi, vii'a tao ra san pham trao ddi la acid.
Su- thay doi mgt so chi ti£u hoa sinh trong san pham len men than cay ngd De danh gia sy thay ddi thanh phin dinh dudng eiia san pham trudc va sau khi len men nudi cay td hgp chiing nim men - vi khuan lactic chimg tdi tien hanh dmh lugng protein thd va thanh phan acid amin.
TT
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
Chi tieu theo lioi
Protein tho (%
idio tuyet doi) Asparagine Glutamine Serine Histidme*
Glycine threonine*
Alanine Arginine*
Tyrosine Valine*
Methiolme*
Phenylalanine*
Isoleucine*
Leucine*
Lysine*
Proline Chii thich ten mau:
Jfir
S y thay doi chi tigu h6a sinh theo
mau JVsl 4,25 0,16 0,82 0,12 0,80 0,15 0,13 0,2 0,2 0,11 0,21 0,08 0,15 0,20 0,05 0,21 0,31
Xai 6,51 0,12 0,91 0,09 0,11 0,31 0,14 0,13 0,25 0,22 0,35 0,15 0,38 0,36 0,21 0,45 0,61 . Nguyen lieu nuoi cay cho pha 1 5 5 %
Jf92 . San pham sau len men ciJa pha
* - acid amin khong tliay the
Ty l§ thay a^i,%
(Jiri2/Jf«i)
+ 53,1 -25,0 + 11,0 -25,0 +3,8 + 100,7 +7,7 -35,0 +25,0 + 100,0 +66,7 +87,5 +153,3 +80,0 +320,0 +114,3 +96,8 dat dp am 2.
Iran Van Chi va Dig Tap chi KHOA HOC & CONG NOnt Kit qua bang 4 eho thay ham lugng protein
tho tang tren 50%. Ro rang da cd sy ehuyen hda sinh bgc tir nguyen lieu thyc vat thanh protein dan bao. Va theo tae gia Tran Thi Thanh: gia tri dinh dudng eua protein vi sinh vat cao hon protein thye vat [4]. De khang djnh lai dilu nay chiing tdi tien hanh djnh lugng thanh phSn acid amin trong cac mau nghien ciru. Kit qua eho thiy chi cd 3 trong sd 16 acid amin dugc djnh lugng la giam so vdi nguyen lieu ban dSu, cdn lai diu tang han rat nhilu, dae biet la cac acid amin khdng thay thi (tang tir khoang 3 din 150%).
KET LUAN
Sir dyng td bgp chung nam men va vi khuan sinh lactic de len men than ciy ngo, ram tuai v^ ed da gdp pban eai thien dang ke gia trj dinh dudng ciia nguyen lieu, md ra mgt hudng mdi trong tan dung ngudn nguyen lieu la phu pham trdng trot phue vu cho muc dieh chan nudi. Sau Ien men pha t sinh khdi nam men tang den 10^ nhung sau pha yIm khi sinh khdi giam cdn lO'. Sinh khdi ciia vi khuan lactic tang din trong 2 pha len men, ket thuc pha 2 sd lugng tl bao vi khuan dat khoang lO'". Qua trinb len men edn la sy
ehuyen hda sinh hge nguyen lieu thyc vat thanh protein don bao, lam tang bam lugng protein len 53,1 %. Han niJa vdi sy chuyin hda sinh hge nay da lam thay ddi thanh phan acid amin trong nguyen lieu, tang ham lugng acid amin khdng tbay the.
TAI LIEU THAM K H A O 1. Blandino A., Al-Aseeri, M.E. Pandiella, S.S.
Cantero, D and Webb C, Cereal based fermented foods and beverages. Food research international 36: 527-543, 2003.
2. Dinh Van Cai, Vien Khoa hpc kT thu^t Nong nghiep Miln Nam, Sir dung phu phSm nong nghiep trong chan nuoi trau bo.
3. Nguyin VSn Mui, An toan sinh hpc, Nxb Giao dye Viet Nam, 2009.
4. TrSn Thj Thanh, CQng nghe vi sinh - Tdi bin lin 3, Nxb Giao dye - 2007
5. Chl Minh Tiing, Probiotic trong chan nuoi, UY -Viet Nam, 2014
6. Dy bao thi trudng dupc phdm dinh du5ng ciia the gidi tilp tuc tSng trudng, Tri thirc v^ phat triln, s6 21, 2005.
7. Tao chl phdm enzyme tai tl hpp thCiy phSn lignocellulose phyc vu san xuat c6n nhifin li^u, De tdi khoa hpc cong ngh^ cSp Nha nude KC - 04 2009-2010.
SUMMARY
STUDY THE FERMENTATION ABILITY OF COMBINATION OF YEAST AND LACTIC ACID BACTERIA BASE ON CORNSTALKS, FRESH STRAW AND GRASS TO PRODUCE THE ANIMAL FEED
Tran Van Chi , Pham Thi Tuyet Mai, Nguyen Xuan Ninh, Dinh Thi Kim Hoa, Nguyen Thi Huong College of Agriculture and Forestry - TNU Using the combination of yeast and lactic acid bacteria fermented cornstalks, fresh straw and grass as animal feed to increase the number of beneficial cell microorganisms supporting for intestinal digestion of herbivores be better. The fermentation was carried out under the two phase - semi- solid and liquid, thereby yeast biomass after aerobic phase increased to IO', but after the anaerobic biomass decreased to 10^. The biomass of lactic acid bacteria increases in all phases of fermentation, bacterial cell count was about lO'" after the 2* phase finished. The fermentation process was the biological conversion of plant material into single-cell protein, increased to 53.1%
protein. Moreover, the biological metabolism has changed amino acid composition of raw materials, increased levels of essential amino acids.
Keywords: yeast, lactic acid bacteria, fermentation
Ngdy nhdn bdi.31/1/2015: Ngdy phdn biin:02/3/2015: Ngdy duyet ddng: 08/5/2015 Phdn bien khoa hoc: TS Nguyin Vdn Duy - Truang Dat hpc Nong Ldm - DHTN
Tei 0965 051219, Email: lranchilna}gmail-Ci 114
