so of ci. Rath, E. Se Va 1, No. (209; 196 386 Shi Sree haters OPTIMIZING THE CONDITIONS OF L-GLUTAMIC ACID RODUCTION BY CORYNE BACTERIUM GLUTAMICUM ARUN SASI, A, KALIDOSS, M, RAVIKUMAR, A.CHANTHRU, TL BHAKYARAJ AND N. YOGANANTH PG. and Research Department of Microbiology, Collegeof Arts and Scence, Puduhkett, Tam Nad, India Key ood: L-Gatn cid Corea gatemicam Bin, Wavy, Optimist [Abetnt- Amino aed sich htc acid were yet hana nique physi and chemi propertas [have many aplication e goa medical and pharracetcal fel Coryebacertm gutaicur me Procuce, age smoune of Loghtams ace hn ther organisms Calture collected form MTCC sti of [rerbol tehsogy. To optimiae the production of Lamu ald by ehanging the carbon source he ‘Kamene lnone plocose ete, manaold, mannoe, sascha sucrose. Temporamue 30. 3- and Ent were ached in 95,65, 710,78 respect oun eee se Aincywilact aan sdcional ad eheapar carbon source. hen esmcetation cf 200 L100 of 1g is hed, Llane ‘ns produced at marian of 30 p/m sets incase ct scarbon source and empectue of 37°C or Frodoction When oun of 100. wou ade 0 ohey the predation wes mui of 275 n/m ith Ecnperntare of 3>C and pol 85 INTRODUCTION GGutamic acid (GA) is one ofthe amino acids that Synthesized in human body. Glutamle 2c often ‘Sats inside all kinds of animal and plans. [thas a Unique physical, chemical of pharmaceutical elds. (Aida et, 1986). Glutamic acid reagents and derivatives are associated vith protein synthesis techniques and research in cell biology and [iochemstry Amino acids compounds destined to become druge polymers and other chemical products, Glutamic acid a intermediate is hlathone one kind ofiver drag. GA happens tobe Se ofthe main components the ell well of Gram= pasitivehaciera. GA is one othe major amino acids Erplan protein and GA plays «role ofthe major ntrogen storage for plants GA has two carbonic [roupe hich ean fom chelates with many’ eet alone, The respectively glutamic acd was used as prescription drugs, that reduced the bod pressure. (Runhwimer et 1970). Coryoebaclerum phtamicum are widely used for the production & L-Glutamme acd. (Kinorbitae 1957) They ae plemorphie, asparogevens, Gram postive bacteria, widely dstoued innature. in lst ‘con of Bergey's manal of ystematic Bacteriology (1969) Corynabacera are involve in 15 section as regular, sporing. Gram positive ods together achiverse collection of 22 genera (Alda eal. (1988) (Qakao ot 1972) this study tried to detect four things i Include, optimieing the temperature, pH, tnd molet important nutrient content (Demein et 1198), Use cheaper medium inode whey obtained form cheese industry ean be used forthe production ‘of L-Ghutamic ac (Peter S-Weaisch ea. 199). (Del Sgunay ef af 2002), And Biotin actas additional Carbon soinrce for L-glutamic acié production (Gletman ot 1992). MATERIAL AND METHODS Corynebacterium glutamicum Is used for the production of L-Gtstamic acd. The eulture was Eollected from IMTECH. Corynebacterium glutamicum has the MTCC number of 26 The calare {Sin the frowze dried form, The revived culture Allowed to grown in Nutrient broth medium and ‘neubated at 30°C for 48 hour. Inoculum was added Jn SOmL of Nutrient broth and then allowed the bacteria to grown, Tn all the experiments, Corynebacterium slutamicum was used. Cols were groven in ———6. Suara. Trlenmayer Flacs on a totaryshacker (150m) at {9°C The following basal medium was used (L) INH) 50, 3g. Urea 5g, KU,PO, 285 kHPO,3H.O 28g, MgS0,7 1,0 0.25¢, FeS0,7H,0, 0.01, MaS0,4H,0 001g, COcl,2H,0, 0.415, ZnSO, 7140, has gH,BO,001g, Coe AH,0 007, Cac GH,0 0.03%, Nici, SH,0 0.0im, Namo 20,24, (im Suga Sg Biotin’ Ig pH (changed) pH-7D. (Clvstan olsenen et a 1950) Different sugar can be used. They ate dextrose lactone, Glucose, {cos Mannie, Maneose Sarh Sucrose, at ancanration 5% Biotin concentration of 200 yg. ‘Tablet Botin 00H) and 100 pg/L was used (Table 1 and Fig.1) {Corian hoishen ea. 1950; Daly, 200, Kimara eal. 1997), In addition to production medium use whey bas the prexiuction medium. 100 mL production medium in 250ml. conical sask with different biotin concentration \08 prapored and sterilized, Dilferent sugars like Dentose, fructose, Glucose, Lactose, mannitol, TPonnoae and sare and sucrose was used 25% of Iovculum was added and incubated at °C and 5° for 48 hours. {Table 2). Fermentation medium ‘dusted pH55, 65,7. and 75. n addon to this ‘they also used os production medium. (Table 3 & Fig 2) without biotin also weed 2s production radia Table 4) SNe Caton Sours Temperature pH cme) | a saa 7 Davos we 16 180 a ut 1 40 130 20 2. Fecteee 2 a ‘uo 0 ve x 83 200 m0 Ee a Glucose #0 100 7 18. m ” 108 2 138 0 4 Lacove » 10 mf ws 180 we 2 = 0 109 5 Mani » 0 rr] 108 09 > 8 1 18 2 6 Mena x 74 Ms 108 128 = Be 168 i 1S 2 Sarch = rat 8 104 rs ” 16 a 0 182 8 Suc » 100 10 ns as 7 m7 1% ra a “Table 2 Biotin (1001) SN Coben Sues Temperawre pH ea a =m are a Py ms 2 2 Faetose 0 03 68 18 are 83 corn) 198 2 Chace 2 2% 20 98 BL y im 2 Ww ma 4 eee > waa 2 iat mz z La x0 22 soa 5 Marit x = wo 10 Bl a & Ww 120 7 6 Mannose x” s2 200 138 180 re at Bo mu ro sort 0 108 = 10 1908; y 0 Bi ct 83. 8 Sucre » sas waka ins we a7 2263 20Optimising the Conditions of -Glutare Aid Reduction by Cory Bates Gitamiaum 187 ear Conor of KGa Ac Rottion by Corye actertum Gstamicam 187 Tables. Whey only SNe Cabos Source Tempera J ee oe ie) Whey oF 10 275 735, me ims 2 bin FY 10s 200 9. 2 ss 3 ets » 109 = vs as 135 0 rr 1 mu ms us pt x use ws mas 105 SS iis “Tae Without stn SX Caren Suse Tenens = Tenperare wy 70 75, rs = TE 5 3 1a a re 240 2 ra 2 acne a os 03m 16 a mM 03 BD ims hos. = no mi ime ” Fe] m2 aa 1m2 4 Lactose » 3 10 im 120 a a4 3 ns 10 5 Manstt » 23 oe 1084 12 y se ime as 12 i » 03 a3. 1103 1082 a m3 os 1203 tesa 7 Sarch x» Se 2 ss 1053 ¥ 2 93 a 1082 % Scere x 105 1 15 rs 7 ins 20 20 to Aller fermentation the total aminoacid content tyasdetermines by Ninhgdrin method of Moene std Stein 1948 with glutamic aid has standard. The calture ites were contrifuged and employed for fstimation of amino acid 2mL of 30% TCA was sulle 010 ml of cultured filtrate and cenit for 20 minates.2 mL of2% Ninhyerin was alded in 2omLov supernatant and heated at OT in weter bath for? minute. The intensity of blue colour then developed wa read at 570 nn RESULTS AND DISCUSSION # temperature, pH, and nuteient _smeentaton forthe production of Lami ac ‘The effect ofcorbon source like Deslese, tricone, slucose,lactoe, mannitol, mannose, starch and ‘Sucrse for Lglatamic and procter. The medium, With pH of 55,65, 7,75 and adaltional natin supplement was biotin, il was used ata concentration of 100 ug/L ‘and 200 pg/L "spectively. Some cause biotin was not used, Whey as alo used asthe production medium, When ‘lots of 200 ig/ Ls sed the L-Clutam acid wos rosuced at maximum of 300 g/mL with ates ‘atbon source and! temperatte of 37°C of pit 6S, Was suitable forbest production. ‘Then dextrose shows maxamm proccion of 23 ug/L temperasure of SAC and pH of 65 when _shicose shows manimum production of 274 g/mL, At temperature of 27°C and pH 65, Mannose sous ‘maximum production of 236 ug/ml at a femperature of 37°C and pH of 65) Slarch shows ‘maximum production of 231 ug/ml ate a femperature of 37°C and pH 6.5, Sucrose shoves ‘maximm production of 23.2 ug/ml at a pH femperature of 37°C and pH 6.5. Manmitel shows maximum production of 149 g/mL at a lemperature of 3°C and pH of 5. Fractve shows maximum production of 138-8 iyg/mt at a temperature of 37°C and pH 6 5. Whey sbowe ‘maxlinum production of 2775 g/L with Blt of 100 ng/L and temperature of 3™C and pH of 65 ‘When without Biotin, dextrose shows shaximam “ip A Producion Meda Withand WiikoutCalice B production of 240 ug/ml a a temperate of 3 tnd pl of 5. |L-latsmic acid produced at axdemum amcun at ‘temperature of 370C and Biotin concentration of 100uL and a pH of 6S and maximom amoun! w: REFERENCES Aida, K/Chitata,K, Nakayama, K. Tabisaa "Yamade, H. 1985. Bjiechnleny of A Pda Elser Vo} 21-87 Via of Spt Batley 1989 ( Iams od Wiking S220 2 CChnitin Hosseten and Reinhard Kramer, 19%0 mate seretion isCo wn games [of Baeridogy 3400-3 DanunceL,Dammadery M, Racha, M. 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