1

I HC THI NGUYN
TRNG I HC KHOA HC

Trn Th Kim Hnh

NGHIN CU S DNG BN THI SINH HC T NC

THI SN XUT BIA NUI CY VI KHUN BACILLUS
THURINGIENSIS SINH C T DIT SU

Chuyn ngnh: Cng ngh sinh hc

M s: 60420201

LUN VN THC S CNG NGH SINH HC

NGI HNG DN KHOA HC

PGS. TS TNG TH CHNH

Thi Nguyn - 2013

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M U
1. t vn
Ngy nay, cng vi s pht trin mnh m ca cc ngnh cng nghip, bn
thi ang tr thnh mt gnh nng cho cc doanh nghip v cc c quan chc nng
khng ch Vit Nam m ngay c cc nc c nn kinh t, khoa hc k thut tin
tin trn th gii. Bn cnh lng bn thi t cc khu cng nghip nng v nh, bn
thi bnh vin. cn c mt lng ln bn thi c thi ra t cc khu cng
nghip ch bin lng thc thc phm.Ti Vit Nam, hu ht bn thi ch yu c
x l bng cch p loi nc, phi kh, sy kh, b hay chn lp, ch mt phn
rt nh c s dng lm phn bn.Qu trnh chn lp, b bn thi v ang gy
ra nhim mi trng nghim trng nh hng n sc khe con ngi.Bn
cnh chi ph x l bn thi i hi mt lng kinh ph ln. Do vic tn dng
bn thi lm nguyn liu cho mt ngnh cng nghip sn xut khc c ngha to
ln v mi trng v kinh t.
Bn thi sinh hc t cc trm x l nc thi ca cc khu cng nghip ch
bin lng thc thc phm c thnh phn ch yu l cc vi sinh vt hnh thnh
trong qu trnh x l arotanh v cc hp cht hu c cao phn t kt lng cng qu
trnh to lng ca h thng x l nc thi do bn thi sinh hc c hm lng
cht hu c c tim nng ti s dng cho cc mc ch khc nhau. c bit, bn
thi t cc c s, nh my sn xut bia c cha hm lng dinh dng cao c th
tn dng lm mi trng thay th mi trng nhn to nui cy vi sinh vt [8].
Vic tn dng thnh phn dinh dng trong bn thi thay th mi trng nhn
to t tin thng c s dng trong qu trnh nui cy vi sinh vt to ra cc sn
phm sinh hc c ch nh ch phm sinh hc ci to t, ch phm thuc tr su sinh
hc, ch phm dit mui sinh hc, ng thi vic tn dng bn thi va gip lm
gim gi thnh sn phm va thn thin vi mi trng. Xut pht t nhng l do
trn, chng ti tin hnh thc hin ti Nghin cu s dng bn sinh hc t
nc thi sn xut bia nui cy vi khun Bacillus thuringiensis sinh c t
dit su.
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2. Mc tiu nghin cu:

Ti s dng cht thi ca ngnh cng nghip ch bin thc phm (bn thi sinh
hc ca nh my sn xut bia) to ra sn phm hu ch phc v cho hot ng sn
xut kinh doanh ca con ngi.
3. Ni dung nghin cu:
-

Nghin cu k thut x l bn sinh hc t trm x l nc thi ca nh my

sn xut bia lm mi trng nui cy vi khun Bacillus thuringiensis.

Nghin cu cc yu t nh hng n kh nng sinh trng v sinh tng hp

protein tinh th ca Bacillus thuringiensis khi s dng bn thi sinh hc t trm
x l nc thi ca nh my sn xut bia lm mi trng nui cy quy m
phng th nghim.

nh gi hiu lc dit su ca dch nui cy vi khun Bacillus thuringiensis

bng phng php sinh hc (bioassays) trong phng th nghim.

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NI DUNG
CHNG I: TNG QUAN TI LIU
1.1. Khi qut v bn thi
1.1.1. nh ngha
Bn c th nh ngha nh sau: Bn l dng cht rn tch ra t cht lng, bn
thng cha mt lng nc ln, c tnh ca bn ph thuc vo c tnh ca cht
lng m n c tch ra[40].
Da vo c tnh ca bn c th chia thnh cc loi bn nh sau: bn thi d
phn hy sinh hc v bn thi kh phn hy sinh hc.
Bn thi d phn hy sinh hc c to ra t qu trnh x l sinh hc (cn
gi l bn sinh hc) hay t nc thi c hm lng hu c cao. Bn d phn hy
sinh hc cng c chia thnh 2 loi: khng nguy hi v nguy hi.Bn thi khng
nguy hi c to ra t qu trnh x l nc cc nh my ch bin lng thc
thc phm, nc thi sinh hot. Bn ny c hm lng cht hu c cao, t cht c
v thun li cho s pht trin ca vi sinh vt. V vy c th s dng lm phn
bn cho cy trng hoc s dng lm nguyn liu cho qu trnh nui cy vi sinh
vt, to ra ngun nng lng, nhin liu c gi tr[15], [17],[24],[26],
[30],[33].Bn thi nguy hi c to ra t h thng nc thi bnh vin, cc khu
nghin cui vi loi bn thi ny phi c x l nghim ngt bng phng
php thiu t trc khi chn, tuyt i khng c tn dng cho mc ch nng
nghip.
Bn thi kh phn hy sinh hc l bn thi cha nhiu hp cht kh phn
hy hay cc cht c. Bn thi kh phn hy sinh hc c chia thnh 2 nhm:
nhm c kh nng x l thng v nhm khng th x l c.Bn thi c kh
nng x l thng p dng phng php thu hi mt s cht sau thiu t, ng
rn to ra sn phm mi phc v con ngi[34].Bn thi khng th x l c l
cc loi bn cha cht phng x v cc cht c d pht tn trong mi trng v
phi x l bng phng php ng rn v chn lp theo quy nh.
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1.1.2.c im ca bn thi
Thnh phn ca bn:
-

Hm lng hp cht v c v hu c cao.

Mt vi sinh vt cao.

Kim loi nng: As, Cd, Zn, Pb, Cu, Ni, Cr

Ha cht hu c tng hp.

Cc cht l lng.

Cc thnh phn khc: ty tng ngnh cng nghip nh cha cc cht

phng x, cht c,
Trong bn thi c cha mt hm lng cht dinh dng c s dng nh
ngun nguyn liu sinh tng hp cc hp cht c hot tnh sinh hc v tng hp
ngun nng lng mi.Hin nay, bn thi c ng dng nhiu khng ch cc
nc pht trin m c nhng nc ang pht trin nhm gim thiu lng bn thi
c thi ra hng triu tn/nm, gp phn bo v mi trng[16],[18].
1.1.3.Khi qut bn thi trn th gii
Trong nhng nm gn y, cc qu trnh x l nc thi vi nhng cng
ngh tin b c p dng nhiu nc hn ch s nhim mi trng t
nc thi sinh hot, nc thi cng nghip.Nhng ch dng li vic x l nc
thi th cha trit v sau qu trnh x l nc thi sn phm ch yu l bn thi,
y l mt trong nhng nguyn nhn gy nhim mi trng.Qu trnh x l nc
thi to ra mt lng ln bn, c tnh chim t 5% n 25% tng th tch nc x
l. Trong qu trnh x l bng cng ngh bn hot tnh, khong 30 - 40% cc cht
hu c c trong nc thi s chuyn sang dng bn hay lng bn sinh ra khi x l
1kg COD trong nc thi l khong 0,3kg n 0,5kg bn. Do , bn thi sau qu
trnh x l nc thi cn c x l v s dng hiu qu.

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i cc nc Chu u, lng bn thi kh trn mt u ngi c thng

k t qu trnh x l nc s cp v th cp l khong 90g/ngy/ngi. Anh, c
khong 30 triu tn bn thi mi nm, tng ng vi 1,2 triu tn bn kh mi
nm. Chi ph cho loi b v x l bn khong 250 triu bng Anh ng vi 5 bng
Anh/u ngi. Sau khi thc hin x l ton b nc thi trong thnh ph ca 15
nc cng ng Chu u vo nm 2005, vic x l ny c th lm pht sinh thm
khong 10,7 triu tn bn kh mi nm v tng khong 38% lng bn. Vic tch
ly ny to ra mt lng ln bn thi[33].

Hnh 1.1.Biu v s gia tng bn thi khi p dng bin php x l nc

thi cc nc cng ng Chu u [33]
Cc thng tin v cc phng php v cc cch tip cn s dng bn thi sau
khi c loi b vn cha c cung cp mt cch r rng. mt vi nc Chu
u, phng php loi b bn ch yu l chn lp t l chim khong 50-75%.Trong
khi , bn thi s dng cho nng nghip nh ngun phn bn ch chim khong
25-35% hoc mt phn nh c ti sinh[39]. Ti Anh, hng nm c khong 18
triu tn bn thi c bn cho nng nghip nh ngun phn hu c, cng nh c
khong 60% lng bn thi ca Hoa K c s dng cho ma mng. Theo ti liu
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ca Hi ng lin minh Chu u (1999 - 2001) c 40% lng bn thi ca cc

nc Chu u c ti s dng li cho nng nghip.
Trung Quc, cc trm x l nc thi to ra khong 5,5 triu tn bn tnh
theo trng lng kh vo nm 2006. Mt phn ng k lng bn ny c s
dng trong nng nghip v phn cn li c chn lp hoc thi b theo cc hnh
thc khc. Trong qu kh, vic thi b bn t h thng x l nc thi c xem
nh khng to ra bt k vn mi trng no v lng bn thi khng nhiu v
vic thi bn khng c quy nh c th. Vic chn lp trong cc bi chn lp
khng ng k thut cng c chp nhn. Nhng hin nay, vic x l bn thi
c kim tra kht khe hn. Trong khi vic chn lp bn thi ti nc ny vn
c xem l la chn c chi ph thp nht th cc n lc v s dng bn thi mt
cch an ton v ch li nh dng cho nng nghip hoc thu hi nng lng vn l
mt hng i mi[32].
Ti Nht Bn, bn thi t cc trm x l nc thi sinh hot s c s dng
ln men k kh thu hi kh metan dng cho pht in, cn bn c dng sn
xut gch Block dng cho lt ng[34]. Tokyo c 13 c s x l nc thi
sinh hot, c t nhiu v tr trong thnh ph x l nc thi sinh hot.
Nhng ch c 3 c s x l lp t h thng x l bn thi, cn cc c s cn li
ch lp t h thng x l nc thi, bn thi s c chuyn theo ng ng
a v cc trm c h thng x l trit bn thi.
1.1.4.Khi qut bn thi ti Vit Nam
Ngy nay cng vi s pht trin ca cc ngnh cng nghip c bit l cng
nghip ch bin thc phm th vn cht thi t cc ngnh ny ang l mt mi
quan tm ln.Ti Vit Nam,i vi ngnh ch bin nng sn, lng thc thc phm
c rt nhiu cc cng trnh nghin cu v cng ngh x l nc thi, nhiu trm
x l nc thi c xy dng v i vo hot ng x l nc cp, nc thi
cho cc nh my sn xut bia, m chnh, ch bin tinh bt, ch bin nng sn, ch
bin thy sn. Tuy nhin, chng ta mi ch tp trung quan tm n vn x l
nc m vn cha c nhiu nghin cu v x l bn thi cho cc trm x l trn.
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Bn thi sau khi x l phn ln c thu gom v chuyn n cc bi chn lp hoc

dng lm phn bn cho nng nghip. Bn cnh trong qu trnh x l nc bng
bn hot tnh c khong 30 - 40% cc cht hu c c chuyn thnh dng bn,
nu khng c bin php x l thch hp s gy ra ti nhim mi trng.
Ti Tp H Ch Minh, tng khi khi lng bn thi c tnh t 3.000
4.000 m3/ngy m (tng ng t 5.000 - 6.000 tn/ngy m). Bn thi cc loi
trn thng x c chi ph thp nht.c tnh chi ph x l cc loi bn trn
khong 300.000ng/tn v trn di 1.000 t ng/nm, thm ch cn cao hn. D
bo n nm 2015 s lng bn thi s tng ln khong 3 triu tn/thng, nm 2020
s khng di 4 triu tn/thng. Trong , bn thi nguy hi hin nay c khong
250 - 300 tn/ngy, cha k n bn thi t cc tnh ln cn a v thnh ph x
l t 150 - 200 tn/ngy[43].Tp H Ch Minh tng thc hin d n xy dng
nh my x l bn Bnh Hng Ha v Bnh Hng nhm mc ch x l bn thi
t nh my x l nc thi sinh hot/ th ti ch thnh phn hu c. Tuy
nhin, cng ngh p dng ti nh my ny vn cha thc s ti u, bn sau khi
x l vn cn rt nng mi v nh hng n mi trng.
Ti H Ni, bn cnh vic x thng bn thi ra cc bi t trng, tnh trng
x cht thi xung cc dng sng cng din ra nghim trng khngkm. Do lng
nc thi sinh hot v nc thi cng nghip x trc tip khng lm lu thng
dng chy, nn cht thi hu c xung sng u lng ti ch, gy nhim, khin
cho c bn con sng T Lch, Kim Ngu, L, St tr nn nhim nghim trng.
Bn cnh , khi tin hnh no vt sng, khi lng bn thi khng l ny li c
trc tip ti cc bi ngoi thnh m cha qua qu trnh loi b cht c hi,
tim n nguy c nhim khng kh, ngun nc Hin nay, bn thi sau khi thu
gom c vn chuyn n b ti cc khu t trng cch xa khu dn c hoc ti
cc ao nui thy sn cn c san lp, thm ch vo bt c khu vc no c th.
Chnh vic bn thi trn lan v hon ton khng c x l nh hin nay s gy
nh hng n mi trng, c bit l tch t cc kim loi gy tnh trng mt v
sinh, mi hi thi. Nghim trng hn, bn thi ang gy ra nhng nh hng nng
n do c b, chn lp khng c lp lt chng thm nn cc cht nhim thm
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xung cc mch nc ngm v nc mt. Vn thiu bi bn thi ti H Ni cng

rt nan gii, hin ti ch c bi rc thi Nam Sn - Sc Sn mi c kh nng x l bn
thi cng nghip. Nu c gii quyt bn thi bng cch tn dng cc bi t trng
bn tm th nguy c gy nhim mi trng rt cao v cng khng c din tch mt
bng ln cha bn thi[42].
Vit Nam, vn qun l v s dng bn thi sinh hc t cc trm x l
nc thi vn cha c cc quy nh c th. Phn ln bn thi t cc trm x l nc
thi c x l bng phng php n gin l sn phi bn.Sau khi bn c lm
kh, gim v trng lng v th tch th s c ng bao v em i chn lp ti
nhng ni quy nh.Mt s t cc cng trnh x l nc thi c cng on x l p
bn bnh.Vi cng ngh ny, bn s c tch nc v p dng bnh. mt s
nh my sn xut thc phm (nh nh my sn xut bia) mt phn bn thi c ti
s dng lm phn bn cho cy trng. Hin ti, vic tip cn vi cc cng ngh x l
bn hin ti nh t hay phn hy ym kh thu hi kh sinh hc cn rt hn ch
nc ta.Ngoi ra cn c mt s cng trnh nghin cu x l bn thi thnh vt liu
xy dng, sn xut gm s, gch lt[4],[6],[12].c bit, c nhng nghin cu
nh gi trin vng x l, ti ch v ng dng bn thi sinh hc ca cc nh my
sn xut thc phm v cc trm x l nc thi lm nguyn liu nui cy vi sinh vt
hu ch sn xut cc sn phm thng mi thn thin mi trng (phn bn vi
sinh, thuc tr su vi sinh) phc v sn xut nng lm nghip. Cho n nay c
nghin cu ca PGS.TS. Nguyn Th Hng Khnh v cc cng s nghin cu x l
bn thi sinh hc lm nguyn liu nui cy cc vi sinh vt hu ch. Tuy nhin kt
qu nghin cu mi ch dng li vic nh gi tim nng s dng bn thi sinh
hc lm nguyn liu nui cy mt s vi sinh vt c ch nh Bacillus thuringiensis,
Rhirobium,[8], [10]. Nhng kt qu nghin cu trn m ra hng i mi y
trin vng trong cng tc x l bn thi mt cch hiu qu, thn thin vi con ngi
v mi trng.
1.2.Phng php x l v s dng bn thi

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10

Ngy nay, trn th gii c nhiu nghin cu x l v tn dng bn thi

s dng vo cc mc ch khc nhau nh: phn bn vi sinh, to ra nng lng
(biogas, in, nhit) hay vt liu xy dng

Hnh1.2. Cc phng php x l bn thi[41]

1.2.1. Phng php chn lp
Trong cc phng php x l cht thi rn, chn lp l phng php ph
bin v n gin nht. Phng php ny p dng rng ri hu ht cc nc trn
th gii. Thc cht, chn lp l phng php lu gi cht thi trong mt bi v c
ph t ln trn. Trc kia bn thi sinh hc thng c x l bng phng php
chn lp, tuy nhin bin php x l ny va lng ph ngun nguyn liu (hm lng
cht hu c trong bn cao), tn din tch ng thi vn l ngun gy nhim mi
trng, nhim tng nc ngm. Ngy nay, cc nh khoa hc ang n lc
nghin cu x l bn thi sinh hc theo hng ti ch BTSH to ra cc sn phm
hu ch, thn thin mi trng phc v cho cc mc ch khc nhau ca con ngi.
Tuy nhin vi bn thi nguy hi th phng php chn lp vn l mt la chn hu
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