Tap chi Khoa hoc-Cong nshfTMy sin _ ".s9-ia'\ . Mimii
-THpNi6,.BAOKH0i
ANH Hl/OfNG CUA CirofNG BO A N H S A N G LEM SINH-rairdNG, HAM LlTCJNG PROTEIN VA LIPID CUA TAO Spirulinaplatensis Geitter, 1925 NU6rTRONGmit>C M4N
EFFECT OF LIGHT INTENSITY ON GROWTH, PROTEtNAND LIPID CONtENTS OF SpiniHna platensis (Geifler, 1925) CULTURED IN SRWATER
Naynlilnfaii: 18/01/2016;NgSyphanbiSnfli&iigqua:03/3/20I6;Ngayduy|tfliB^15/&2D16
T6MTAT * Thi nghiim duoc Ihtrc hien di danh gii inh hiring cua cuing diinhsangflif sinh miing,ti,nll0li
protein vi lipid cua tao Spirulina platensis. Tia duoc nuoi trong binh 500ml vdi 0,m$n 30- 32%o, 65 mic cuang0 inh sing: 1.000. 2.000. 3.000. 4.000 va 5.000 lux. Kel qui cho thiy: Q,dng0inh singinh hutng len sink truing oia tia. & 1.000. 2.000. 3.000. 4,000 vi 5.000 lux dot mil 0 cifc ddi lin Und li 1,2) ODt 1.36 OD: 1.64 OD: 0.95 OD vi 0.84 OD (F<0.05), oxmg 0 mifc 3000 lux dtH mit0 Clfc dql cao nhii.oh^
nuoi Ihi 13. Cu&ng do inh sing cung inh huong len him lupng protein vi lipid cua tia, i 1000 lux him,limg protein vi lipid lan bm 0151,09% vi 8,28%. a 2000 lux (01 60.14% vi 10.05%), 0 . cao nhat a JOOSj/ljt (67,78% vi 13.15%). 4000lux (0145.22%: 6.81%) vi5000 lux (0138,71%: 5.65%) (P<a,05). Til0,^0
nuoi tio S. platensis trong nuoc man a cuing 0 inh sang 3000 lux li Ihich hpp. '7, Tu khoa: Cuang do inh sing, ham lugmg protein vi lipid, mai 0 cifc dai, sinh truing, too SptnfUna
platensis ABSTRACT
This research was conducted in order lo esdmale the effect ofUgld tntemlly on growth, prceln and lipid canlenls of Spirulina platensis. Algae were cultured In Erienmeyerflask in volume of500ml In 5. levels of 1.000. 2,000. 3.000 and 5.000 lux wUh Ihe salinity between 30-32%,. The results showed d«,l, llghtMeml^
effected on Ihe growth afS. platensis. In which. 1,000. 2,000. 3.000. 4.000 and 5.000 lux had die maximum densily afl.23 OD. 1.36 OO. 2.64. 0.95 and 0.84 OD. respectively (P<O.OS). Specifically 3000 hagme the highesi maximum density on the day 13 Similarly, light inlenslty also effected on the protein and lipid camene ofS. platensis. In 1000 lax. Ihepmein and lipid canlenls of algae cells wen 51.09% and 8.28%. in 2000 lia these value were 60.14% and 10.05%. in 3000 lux Ihe highest results were 67.78% and 13.15% (P<0,05), Meanwhile, the protein and lipid contents afS. platensis cultured in 4000 and 5000 lux were 45.22%. « ' « ; » and 38 71%. 5.65%, In summary, il can be cultured S. platensis in seawater at 3.000 hoc was the most suitaUe light intensity.
Keywords: Growth, light inlenslty. maximum opdcol density, protein and lipid cements, SUruUna platensis
' Vi#n Nuoi trbng 9\iiy sSn - Tntimg Dg, iigc Nha Trang
124 . TRUONG OAI H O C NHA TRANG
Ti^tAi
S l * » * i a laioai l4o lam i f t f l B u d M i B u S i *
" * " * " '"WIS piotein c h f e r w s e ^ T T K H A l W n « » * . 9»u vtortn; («A, khdang, axil a n i « i v a c a c a x l l b a o t h * j ^ n , B & ! C 9 n h a o , Wa nana thfch ling ^ t v a i ylu l l m a s u f t n g , I aeukWnvakylhultnudlaonjaneflnglamat
Irong nhOng lgi t h i tnong null sinh khil loai lao
* W . Do aa, B o Spinillna da auuo nghian
• * . sSn xult va ling dgng Irong nhilu ITah vvc Ciia aai slngnam Ihifc phim chiic nSng I- w a n dinh du©ng b l sung thilt ylu, thu&i
* 0 a t)$nh(ung«>u., HIV/AIDS. vi§m gan, lHu
<l»ft>ia,...), m f phim (cham sftc da va tac), Ihiic an Chan nuai va xtiiy nufto thai (IJ.
T * ) Spirulina nuai trong mat tn/ftng nuftc mSn c6 gia tij dinh duftng v.«7t u^i han so vai nul) .fcong jiuftc ngai t h i hi$n ft s l lupng chit OS hoat tinh sinh hpc cao (polysaccharides.
Inositol va phycocyanin), cac nguyin 16 vi
!»7na, ham luftng protein, lipid, cac axB bao thill ylu (DMA, EPA, ARA. LA, LOA,...) [10J.
N ^ l gia tri dinh duftng cao, null tao Spirulina trong n»»o mjn can gap phln sit kiam m|t Iwns Iftn cac chit khoang da lu(/ng, vl luvng M sung va isn dyng lit tilm nang dian Hch i1«Sc mJn sin c6 6 nufto ta. Tuy nhlSn, chira BiSlifcM cang Wnh nghlln cOij nao ft nuftc la
* o«p din vl|c nuai tio Spinjiina trong nuftc min. Chinh vl viy. nghlln oiiu dupe thire hiln n,»lm xlc ainh cuftng d | anh sang thich hap oho, nuai tao Splmllna platensis trong nuftc g n ft aHu WSn Ihl nghlSm, la oo sft a l thilt
* quy Irlnh d n g ngh$ null sinh khli loai tao
•«y *Ouy me d n g nghllp phuc vu nhu d u th|
fMrng Irong nuftc va xult khiu.
* Anh sing la nguin nang luyng chinh cho qua With quang hap, gap phln vao vl|c san X * ngoln sinh khli cho vi tao. Trong aa, W n g a? Inh sang la ylu 14 d Snh huftng trvti
•Sp a i n cuftng aa guang hap ofia tao nhit la
So 2/2016 tengailulii&i mgtaa cao [11], Theo Uvens « Soisalooa(1996) [12] cuftng aa anh sang thich hap cho hiu hit cac toai tao nim trong khoang
» 2.500-5.(KIO lux 6 quy m l phing Ihl nghiem va tft 20-30 Mux ft quy mo ngoai tifti. Viec xac
* * cuftng ae sing thich hap cho ttmg laai tio. mat ae ban diu. t h i tfch null va dp sSu ling nuftc nhim nSng cao nang suit. Bit ki0m duoc nguin nang luong dien va rtH ngan thfti gian nuai, dem iai hlftu quS kinh t l cho nguai null [8,14]. Tuy nhiSn. cho ain nay cac nghiin Ciiu v l anh huftng cia cuftng aft anh sang ISn sinh Iniftng cua tio Splmllna trong mil tniftng nuftc man vin chua h i auac a l cjp. NghiSn ciiunaynhimxacainhcuftngaaanh sang t l i uu cho nuli tao Spiiuiina nuftc mjn trong dilu ki»n thi nghijm, la m i l trong nhDng Gin a l d l thiit lap quy trtnh d n g nghj null sinh khli ioai t«> nay ft quy ma ceng nghijp phuc vg nhu d u cho con ngufti.
ILVATUEUVAPHlTONGPHAPNGHieNCOu I . V j t I l j u nghlln cOu
Oil tuang nghiin cCm la tao SpmMna platensis dup^ thu thjp tCi Hr nhijn, d nguin glc nuftc ngpt (2%.) va aa airac thuln hia trong mii inrang nuftc mjn (30 . 32%,).
Nghlln a i u duac thirc hijn t;l Phang Vi llo Ihujc Vijn Null liing thiy sin, Tmtmg 6,1 hoc Nha Trang.
Nguin nuftc biln sii dung cho nghiSn a i u nay duac xii ly bing Chlorine 25 ppm trong 2 ngay, sau d l phcrt nlng va trong haa bing NalHIhkjsulfate vai t} n 1 : l . Tiuftc khi thi nghijm, cjc binh Ihijy Snh vfti Bil Uch 500 ml d u w dua vaa hip Sjt Sling ft 121'C, 1 aSh song 15 phlit.
2 . B i H t h l n g h l j m
NghiSn a i u jnh huftng oia cuftng d j jnh sang anrc sin hjnh ft 5 miic cuftng aft anh sang khac nhau gIm: 1.000, 2.000 3 000 4.000 va 5.000 lux. Anh sang auyc b l irl bing'
n t t t N G DAI HOC NHA TRANG . 125
Tap chi Khoa hgc - Cong ngh4 ThSy ajg. S&2/2&16
cac b6ng Shn h u ^ h quang, anh s^ng b]ing,Gd c6ng su^t ti> 30 - 60W. Tao giong s& dgng ptra M nghiem dang o* giai do^n tang tm-o^g ( c ^ pha iogarit) v6i m#t dp ban &k\x IS 0,4 OD difoc nudi trong moi taramg dinti dtfdng V2 (Guillard, 1975). Moi nghi&n thOc dupe thytj hien win 3
\hn l^p cCing thdi diem.
CSc y6u to mdi tn/ong dipcrc duy tri bong pham vi thfch hap v6i sinh tnrdng cua teo S.
platensis: nhifit d0 phong 25°C, pH: 8, ch^ dO chieu sang 16 gicr sar^: 8 gicr t6i, SLIC Ifhi lien bfc. dg m^n 30%o.
3. Phipomg phdp thu t h | p vd xie fy s6 l i f u 3 . 1 Phircmg phap xac djnh sinh tnrong cUa tSo
Theo Coutteau (1996) [9]: Sinh tnj&ng ci]|a tdo duvc xac djnh fhfing qua m$t dQ quang (optical density-OD), bang c i c h l5y 5 ml dung dich tSo dva v t o mdy do quang pho (Spectrophotometer CECIUCE 1011, 101S, 120554) d bu^c sdng 560 nm.
3.2. Phuang phdp phSn tich hdm luring protein vd lipid
D S phan tfch Idm lupng prot«n vd lipid, cdc mSu tdo duvc thu t^i th&i diSm cu6i pha tdng trirdTig (pha Iogarit), khi tdo dgt sinh kh6i ci/c dgii. Cdc mSu tdo d u ^ c tdch ly tdm kh6i mdi trud'ng nufii b i n g mdy ly tdm oS t6c dfi quay 7.500 vfing/phdt bong thd'i gian 10 phOt, sO- dyng mdy iy tdm Hereaus Suprafuge 22. MSu t5o SSU dfi dupe s^y khfi & nhi$t dfi 55'C trang 2 gid-, luu giu d- nhi$t dfi am 20°C tofdc khi thyc hi$n cdc phSn tic^ hda sinh. Hdm lupng lipid trong mSu tdo Kt\6 dupe phdn tfch theo phuong phdp cua Bligh vd Dyer (1959) [7] sir dgng h5n hpp tdch c^i^t chi>a chlorofonn vd methanol vPi tJ l | thS tfch Id 2 : 1 . Khodng 120 ml cua h6n hpp ndy dupe si> dgng dS tdch t^ik m6i gam tdo khfi. Cdc ch^t r i n dupe phdn td(^
bang cdch SCP dgng gi^y Ipc rfiuySn dgng 2 Idp ciJa Nhat Bdn. Cdc eh^t hfia tan vd dung mfii dupe Idm bay hcri trong mdy hiM chdn Wiflng
126 • T1?LJ<!)NG OAI HQC NHA TRANG
d hht$t dO eO'C. Qud ftlnh tdch tchldt A f p c ttm^h!^ 3 lltn d)o dSn khi thu dupc todn bO lupng fipid thO tror^ mSu tdo. Protein thd eugc phSn tfch b4ng ptuicmg phdp l^eldahl cSa AOAC (1998) E51. Qud trinh phdn tfch dupe ti4n hdnh tgi Vi$n Cdng n ^ $ sinh hpc vd MSi tru6ng - T m ^ n g Dg4 h ^ Nha Trang, 'i 3.3. Qudn If cdc yiu M mdi frudng
Cdc y^u t6 mfil tiVdrng dupe xAc ^ h 2 ngdy/lan. Nhi^t dfi do t ^ n g nht^t k4 nfpu (d9 (*[nh xdc 1°C). pH dupe do bang mdy flb pH (Manna pH Meter, d0 chfnh xdc 0.1). Sfi m$n dupe do b i n g kh6c xg k^ (ReftactomelH', dfi chrnh xdc 0,5%D). Cud>ng dfi dnh sdng A f p c d o bang mdy do Hanna D l g ^ l lllumlnometw-D^i Loan (dfi chfnh xdc ± 6% gid trj a?c dupe).
3.4. Phuvng ph&p xO- /j? s^ li$tj
Cdc sd li$u sau KhA thu t h ^ dupe p h ^ tfch bSng phdp phdn tfch phucng sai mOt y6u t6 (ANOVA) tr«n phSn mSm SPSS 16.0.1^1 c6 sp khdc biat giua cdc gid trj trung blnfi' v^ m^t dfi cpc dgi hay hdm lupng protein vd lipid cQa cdc nghi#m thil-c, phdp kiim dinh Duncan's Test dupe s * dgng 6k xdc ^ n h sv khdc bi^t c6 'jf nghTa th6ng 1 ^ vd-i mi>c # nghTa P < 0,05. rk cd s6 lifiu trong thf nghigm 6\m trinh bdy dufri dgng Trung blhh (Mean,) i. Sai s6 chuin (SE).
III. K^T QUA NGHIEN CCTU VA T H A O L U p ^ 1. Sinh t n r d n g cda tdo
Kk qud cho t h ^ , cutmg^ d$ dph. S ^ T E | dnh hudng r5 r©t d i n sinh t n ^ ^ , t p ^ d | ^ dgi vd thPi gian dgt m f t dO cg-e i ^ i ciia l^o S. platensis. v * i xu hudng ehung. Id C3i|^.
dfi dnh sdng d- mi>c tiilp t6t hon.fr mfi^cdii;
0 | c bi§t ^ o difpc nufii & cudng dO Snti ^ ^ 3.000 lux cho mat ( ^ cpc agl IPn n h ^ f i n ^ l nMfii thfr 13 va khdc bl$t so vPi cdcmite edtiil,'^.
(p<o.m}ih\m'i). 5 |
S62/20h
" » 21 23 9 II U B
» « eta (new
T i a aui^c n u l l 4 cuftng aft anh sang tang
* B . 1.000,2.000 va 3.000 lux cho mat aa 'VC aai tang a i n (1,23 ± 0,03 O D ; 1,36 t 0 02 OD va 1.64 ± 0.03 OD)i ft cac mftc caa (4 000
ft95 ± 0,04 OD va 0,84 ± 0,03 OD) (P<0 05)
W . hu^ng a i n ftai gian duy M q u i n B i l d a Sa. Tnmg aa, tao auffc n u l l ft cuftng aft anh y n g f l i l p (1.000,2.000 va 3.000 lux) Sifti gian I ^ W q u I n B i l l a 2 3 - 2 5 n g a y savft!nhftm ca c « n g aa anh sang cao (4.000 va 5.000 lux) fcftl gian duy tri quin B i l la 15 ngay (P<0,05)
cac quan sat fliSm cftn cha B i l y t i o S. fltetensis dugrc nuai ft cac mftc cuftng aft I n h ^ n g caa (4.000 va 5.000 lux) eft mau s i c W bUn a l l Sl. mau xanh dam chuyin sang
C ^ aa anh sang caa can gSy ra hiftn tugng
« c d ^ bam v i o thanh binh thanh tftng mang I " ngay nuOi Biu 2 Sft di cho a i n khi tan Hii
Cuftng aa anh sang augx; xem la y l u t l quyit ainh d i n cuftng aft va sSc aft quang h ™ cua Ua. Trong giai han nguftng, cuang da i n h
^ n g «la Biuln vfti cuftng aa quang hap-sai lai nlu vuat qua nguftng Uil cuftng dft anh sang l a i » I I nghjch vfti cuftng aft quang hap [15J
Khi cuftng aa anh sang tang se ttiiic d l y qua trinh quang hap va phan chia t l bao diln ra manh me. k i t qua la sinh khli tao tang iSn nhanh Chang [14J. D i l u nay 11 giai 1,1 saa Sang
t u 1.mo - 3.000 lux Cha m|l d | cue dai tang aan va aai caa nhit a nguftng t l i uu 3000 lux W lai, mftc cuftng a« anh sang cao 4.000^
5 000 lux la nguyin nhan lam ftc c h i qua Wnh quang hyp va Saa a l l chit trang t l bSo
»i_am chi lam chit t l bao. Nghiin cuu ciia T i n Nu My Nga (2006) [2J san tao Chaatocams sracfc va Phan van Xuan (2010) [3) ft tao TViafess/asra sp. cung cho thiy, nguftng anh sang 111 uu cha 2 ioai tao nay nam trong khoang tu 3.000-3.500 lux.
2. Ham lirpng protein va lipid c i a t i e Cuftng d l anh sang ca anh huftng Iftn d i n ham luang protein va lipid d a tao S. p/a(ens/s
("hinli^"?.T"''"*^*'"'*""""9''*'"
(hinh 2). Tao duprc null trang dilu kian cuftng
^ ' ^nh sang ftip (1.000 - 3.000 lux) cho ham lua^ng protein va lipid caa hun sa vai null trong ft^cuft^ngaa anh sang cao (4.000-5.000 lux' Ham luwig protein va lipid trong t | baa t^ng nhanh B « „ cac muc cuang da anh sang
TRUONG OAi HOC NHA TRANG . 1 2 7
Tgp chi Khoa hoc - Cone nsfle Thiy sin
1.000 iux (51.09 ± 1,12% va 8,28 ± 0,58%), 2000 lux (60.14 ± 2.45% va 10,06 ± 0,65%) va dat cao nhatft 3000 iux (67,78±1.08%va 13,15
± 0,94%). Tao duac nuai ft mijc cuftng dQ anh
Si 2/21116
sangfcaa pdOD va $.0)0 kix) cho h i m luang prataintVltiplttlch 1% Irong ^ baa H o la lhi|>
nhll,. l l n luftt la 45i22±2,13%; 6,81 ±0,88% va 38,71 ± 1,88%; 5,65*0,72%) fP<0,05;.
• U p M
IJMOtax 2 j n 0 l u x 3.omtax 4 M 0 k l x S.OMIgx CtrdngOlanhsaiM)
Htob 2. Anil hii«iig ciaciremgllSinhsingBnliiiiilinmBproleliivillpldclintilo Cdckynra.b.c khac nhau tren cic c$t Ihi hiin su khac biit thing ke (P<0.05) Thfti gian chilu sang, cuftng di) vi chit IV. K6T LU*N VA Kltti NGH|
luyng anh sang la 3 Bianh tl quyit ajnh din qua trinh sinh tnfftng va phat triln ciia cac sinh v i t quang hap nai chung va cac loai tao n i l n'Jng. Trong aft, cuftng dft anh sang duac xem la y l u t l d i u liSn anh huftng true tilp a i n su l i n g hap cac thanh phin trong t l bao [13J.
Nhilu nghiSn cftu dS chi ra ring: s v t i n g hap protein, lipid va carbonhydral ft t l bao tao diln ra manh mg n h l l Bting d i l u ki$n cuftng dft anh sang dsl nguang t l i uu. Nguac lai. su l i n g hap cac chit nay s§ b| dinh tra trang tnrftng hyp cuftng d l anh sang vuat qua nguftng, ICiy theo laai tao, m#t dft nuOi, B i l tich nuai va aft sau l i n g nuftc [8, 9,14].
Nhu v j y Irong nghian cftu nay, cuftng aft anh sang 3,000 iux la mftc ttiich hgp cho tao S. platensis nufti trang nuftc m j n 6 a i l u kiln thi nghiim nham dat duac cac ttilng s l t l i uu v l sinh t m * n g , mat dft cue dai, ham luang protein va lipid.
1. K i t lu$n
Tac dua'c null ft cuftng di anh slnjti*
1.000, 2.000 va 3.000 lux eho mjl Si ocrc a«l tang d i n (1,23 ± 0,03 OD; 1,36 ± 0,02.013 vS 1,64 ± 0,03 OD) va dat cao n h l l ft cuftng i ( ! 3.000 lux ft ngay nuOi IhiJ 13 (P<0,06). Clfftis d | anh sang 4.000 va 5.000 lux che m j t d?
eve dai r i t t h i p (0.95 ± 0,04 OD va 0,84 ±0;0J OD) (P<0,OS). Thai gian duy tri quin BlS oia;
tao ft mftc 1.000 - 3.000 lux la 23 - 2S h f f ^ dai han so vfti mftc 4.000 - 5.000 lux (15 ngaj^
(p<o,o5;.
Ham luang protein va lipid tieh iOy Bang t l bao tang d i n theo cuftng dft anh s i n g 1.000 lux (61,09 ± 1,12% va 8,28 i 0,58%), 2 . ( i 9 lux (60,14 ± 2.45% va 10,05 ± 0,65%) v l d j j cao n h l l ft 3.000 iux (67,78 ± 1,08% v l 13,16 i 0,94%) (p<o,05). Mftc cuftng dft anh airtg eao 4.000 va 5.000 lux cha ham luang proW) va lipid r i t Blip, l l n luai la 45,22 ± 2,13%;
6,81 ± 0,88% va 38.71 ± 1,88%; 6.65 ± 0 , 7 2 « ' (P<0,OS).
128 . T R I / O N G OAI HOC NHA TRANG
Z. M i n nghj So 2/2016
apfctensfenuSitrong nuac man. ' ' ' " " ' " ^ "^ "^ '>*"^y< <^ trong t l bao tao
X A l L ^ T H A l a K B A o
2. Ta« NO MJ Nga, 2006. NghiSn cim 4 J , h , ^ ^ ^ ' " ' " ' • ^ " " " S B s . h o c C & T l i a . T r . 1 5 - 2 7
lAlgAnh: ^
:'• T^:.v^:f«->-M»-»-p™-..or:x:rrr^^^^^
r ; ' C o q x , n > d o ^ l ^ . r s S . 2 9 - 1 0 " ' ' ' • ' " " " " " * " '"r iceiing ™™c i„v,:n.|„«es. Plen,„ p„t„,,i„^
: W R r a a a i S. 1«L, Pury D. L., h. V Thinh KLO C Pri
I " T ^ " " - ^•»™l of Applied Phycology. KluwrAcad J L S t o "^ " ^ ^ ^ « " ' « ' " f" u,e ,„ „a,Z fS. U. K. Ch.uh» „ d Neemj P.ihiU (2010). EffM of differen, ^ ^
* » » / » , W » ™ . J „ ™ | „fAlg.| B i „ „ , ^ , I (4), g ™ ° 7 ' " ' ° * ° ™ ™ *'= Pnxiuciio. of chl„„pVi| ^y 16. Miijshrt A.. K«,cb.„Jcs. N., Bimmg B. ^ Tmdchmcp M (19961 n , „ •
i>=cl™„on process of the cy»ohac.eri™ Spirulina platensisLX^^i^ " ' " " " " y " " ' - «" fe ^
"'"••••" 1
W J O N G OAI HOC NHA TRANG.
