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distill.d wlter. Tle conlcnts wc frozen with dry ice dd tet tube w6 evacuated<br />
with ihe helt ofa slclion punp. Thc tube wd Bealed with pin poinl oxy8en suigas<br />
flame and placed in uprigil posilion in an ovcn reeulaled ar I looc for 22 houc.<br />
'fhe hydrolyate so folred ws evapoEred unde. vaculn 10 d.ynes ro reDove<br />
Ilcl. TIE d.ied nln of hydrolrate was susF ded witb 2rnl buifer (rrH 2.2). Tle<br />
solublc maicrial was ccnhiluged to get rid of visiblc sedihen$ sd samplc wd<br />
loaded on ion exchdS€ column of aubmatic miro acid analy&r for g€ring<br />
compl.le profile of mino acids lr€sent ir the smple. Standard solulion of rhino<br />
acids qaalsorun in lhe aminoacid analyzer under simila. conditions.<br />
6, Ribos€ Nucleic Acid (RNA) Estimation<br />
Tlt€ co.centralion of RNA5 in th€ fermenr€d biomass was €stim.led<br />
by rh€ method ofCerioui(1955) as Dodifi€d by Munro and Fleck (1966). For fiis<br />
purpose a stand{d cupe Nas conlrrucred by rakin8 different votumes ofsranddd<br />
dbose solution (0.01% in concentrared HCt) in six rest tubes. whereas the 7th lube<br />
ser.d as blank (Table- l2). '[re direrenr dbose corccntarions as well as th. blank<br />
werc lrealed *ith lhe rcagenls as illustmcd in rhe rabtc.<br />
TIle test rubes shaken rhorouShty were immersed nr boiling watef bath<br />
for 40 minutes and tlen cool€d undef tap water. Tte cotou| was rhen exracled {i1h<br />
5mL anyl alcohal by mixing md c€nlifu8ing ar 4000 rpm for five minutes. Tle<br />
extracted blue colour€d solution was @nlrituged again ed iB optiql densily was<br />
Md on spectrophoromeler ar 675 nn againsl rhe blank. The stmdard cune so<br />
prepared is presenr€d in Fi8,5.<br />
finely ground and seiv€d lhroud 40 mesh sMpl€ of fermenred<br />
biomais (0.1 g) was exkacred rwic€ with chillcd absolure alcrhol (5ml each iime)<br />
lhen centrifuged for five minules al 4000 rpm. The residue was once exracred wilh<br />
chilled l0 percent Ticholoro-ac.ric acid (TCA) by cenkifugation and rhe<br />
supemalrnl wd discard€d. l.lle r€sidue was rhen fina y treal€d with 5 peoent TCA
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