‘ese url of Beil Scenes Taorescece Pet 3, an “Mc 209, p.168-177 Effective Production of Biomass of Aspergillus sp Using Different Oilcakes “Arun Sasi, R. Bhakyaraj, N. Yogananth', A. Chanthuru! and M. Ravikumar? "Deprnet of micobiolce, JJ. College of rs ane Science, Pedestal 622-422, ‘Deparment of Pla Biology and Bhtechuxogy, Cant. Are Coleg ee Mest ‘Naaman, Chern “Corresponding author: varamicro® gmail.com ReceWed October 29, 2008: reviewed January 1, 2008 accepted laniry 28,2009 Abstract Grom of Apergitaswpecis on arcu wast aera ike ronan icc, Sesame, ont cake medium in submerged fermertaton were used to determining the dy Wowie, eae pra prota ‘Key words: Biomass. Aiperyilu sp, Oileakes, Marte sponge. Submerged fermentation Introduction Marine fungi have the ability to grow at certain sea Water concentrations Gobrson and Spacrow 1461). They Ae mostly facultative fungi (Abbot, 1993) and can be solated from marine sponges (Ainsworth, al, 1973). Among these fungi A niger and A. oryzae were selected ir the present study: Most developing counties ofthe ‘orld have been facing the problem of malnutrition due © the deficleney of protein i human food snl sno ed, duct the rapid growth of population. is therefore, "portant to nerease protein prvkiction by utilizing all available waysand means tomect the increasing word mani fr protein rich food. This led to the search fora 18 non-conventional protein source. A great deal of interest ‘hasten focused on the potential for syrheszing miceobia Protein fom agricultural waste lke ground nut ilcake, coconut ilcake, Sesamum oilcake. (Ravinder Rodravaram and Anujkumar Chandel, 2005) . These cileakes were used as ingredients inthe preparation of fermentation media (Hirokilshide etal, 2004, Oshome ‘nd Lkenebome, 2005 and Heidelberg and Speingee Bein, 2006) in the present study. In adition to this he stfect of Aifereot physicochemical parameters, lke the effect of ‘Various nitrogen sources and eatbon sources on biowsss Production and total protein content were also sti bse a So fonlaf Reseach Journal of Blologkcal Sciences Ienorescence Foret 3, an ~ Mat 2009, 1p. 168-177 fective Production of ioms of Aspergillus Using Different Oieakes Cileakes are byproducts obtained after cil extraction, The chemical composition of cilcake varies depending upon the extraction metheds. These oil cakes are fitly rich in protein and are used as fodder for farm animals and as agriculture fertilizers (Singh et a, 2003). They are also sed in the area of enzyme fermentation for utilization of raw materials for the production of value-added fine products (Pandey eral, 2000). Oileakes ate ich in fibre and high concentration of non-starch polysaccharide. Coconut cileakes contain 14-20% of erude protein. Ground nut cilcake contains 40-50% of crude protein. Generally oitcakes have less fat content which ranges from 2:3% fat, This study indicates that cileakes are ‘cheaper for preparing production media when compared to commercially available media and that we can produce more biomass and single cell protein using these media ‘than through the normal media Materials and Methods Isolation and Maintenance of Fungal Culture Marine fungal strains like A. niger and A. oryzne were used in the present study (Ainsworth ef al, 1968), ‘These organisms were isolated from marine sponges (Angitae gece al., 1998). The two species of Asmereillus ‘were grown in potato dextrose agar medium for seven days at °C and then stored at 4°C (Abbott 1993), Sporulation Method ‘The sporesof the fungal strains were added tobroth stored at 4°C (Abbott 1993)-Ten ml of distilled water containing 0.1 % of tween80 was for seven days a 10 transfered for sporulation (7 days old PDA culture) “The spores were dislodged using the inoculation needle under aseptic conditions and a suspension of appropriate solutions was used as inoculum (Pandey, 1994). Production of Fungal Biomass Different oileakes like groundnut oil (GOC), Sesamum cileake (SOC) and coconut oileake (COC) were used as substrates, Al eleakes were procured from the local market in Pudukketta, Tamilnadu, India. The submerged fermentation method for culturing the Fungi was carried out in shake flasks. Sgms of powered dry oileake was taken into 250 mi Erlenmyer flasks and supplemented with (NH), SO, (2.0 gmf), KH,PO,(.0 a), MgSO, 7H, O(05 gf) and ZnSO, 7H,0 (0.1 2/0. Addition of cstiled water was adjusted vo get the required ‘moisture level. Then the media were again supplemented with different carbon sources like dextrose (2.0g/1), glucose (2.0g/1) and lactose (2.0g/1) (Sumitra Ramachandran er af, 2004), For studying the effect of various nitrogen seurces in biomass production the media were supplemented with NH,Cl, NaNO, KNO,, Peptone, Ure, Yeast extract and (NH4),SO, at a concentration of 1.7, 25, 1.3, 1.6, 24, 3.1 41.2 g/t we and Lkenehome, 2005). The pH as maintained at 3.5 10 8.0 and the temperatire of the respectively (Ost media was adjusted to 20 10 40°C. The culture flasks were placed on a shaker for nine days at 120 rpm (Pangpei Khang and Ibrahim Ohemer 2005). “The growth medium was pasteurized at 65°C for 30 minutes in a water bath after each fermentation period‘Research Joumal of Biologia Sclences Taflrescence I, Hort 3Jan- Mat 200, pp. 168.177 Mycelia were removed from the flasks by passing through a dried and pre-weighed Whatmann No.1 filter ed water. The filter paper was dried at 90°C — 100°C by using hot air ‘oven, Then dry weights were obtained (Elisasliveli 1993) and the protein content of the dry biomass was obtained by using Lowery method. The results were tabulated, paper and washed twice with sterile Results and Discussion ‘The two species of fungi A niger and A. oryzae were isolated from marine sponges and empleyed for their efficiency in the biomass and protein production “This study was done by growing these two fungal species in three different oileake medi supplemented with different carbon sources inthe form of dextrose, glucose, lactose and different nitrogen sources in the form of NaNO, (NH),SO, NH,Cl, KNO,, peptone, ures and ‘yeast extract Both the species responded well when grown in {groundnut oil cake medium supplemented with the above joned carbon sources and nitrogen sources. The highest protein and biomass productions were recorded in glucose and NaNO, supplemented medium, which sere 0.9640.0 ig/ml and 3.060.03g/ml respectively for A. niger and 0.9020.02ig/ml and 3.0340.01 g/ml respectively for A. oryzae (Table 2 and 3) ‘When nitrogen source alone is supplemented both species showed comparatively less diy biomass production than in protein and carbon supplementation (Table 3). When A. niger was grown in NaNO, supplemented medium the biomass production recorded ‘was 1,06 £0.01 g/l and the protein content recorded was 0.92 + 0.01ig/ml. For A. oryzae these were found to be 0.08 40.001 g/1 and 0.824+0.02ig/ml respectively (Table1, Fig), ‘Table 1. Effect of Groundnut Oileake Medium Supplemented with Different Nitrogen Sources, ‘on the Biomass Production and Protein Content of A. niger an A, oryzae [Oran used Ritopen Source [Toil Prin Diy Biomass ig) (git) A niger | NaNOy ‘092001 062001 (NHL): 50. 063 £001 {00 = 0.03 f NHCI 0320.01 096 +0.01 KNO, Os 2001 on3 002 Peprone 0440.03 o712001 urea 0320.01 056003 Yeast extract 046 = 003 0640.02 A. omzae | NaNO} 082 = 0.02 0882001 (NH); SO, 0.72 £0.02 085 + 0.02 | NHI 053003 0722001 KN, 063003 0632003 Pepione 0432003 070+ 0.04 Urea 0642001 0620.02 ‘Yeast extract 0480.01 053 -Research Joural of Bilogical Sciences Inioresence 1 Faet 3, Jan Mar 2009. pp 168-177 ifecuve Production of Biomass of Agpergis p Using Different Chen Figure 1. Effect of Groundnut Oilcake Medium Supplemented with Different Nitrogen Sources, eee ‘on the Biomass Production and Protein Content of A. niger and A. oryzae sis as Aor aA se The : ‘ecorded ead vit 2 | vel or oe nea Demi | Wesieazy Cet | Pup y — ro eas ‘able 2. Ete of Ground! Oieake Medium Supple wit Differ Carbon and oe ‘Ntrgen Source, onthe Biomase rosueon and Pein Geren 7 ‘Orraniion | Carton | Niwogen | Toual Proacla ‘Dry Biomass vail vom | aoe | nome | ue i [>on] ma} oot: a Seer | mora || -emaeer | geen be Lactose NaNO, oneaor 262x001 ablel, Dextrme | HDSO | 0.632001 108 +002 fener | omen | amaenr | terrane tmie | oatoo. | asszeot | tarsoun diiomee|| naan | sumsane | optctee fee | msc | esscean | concent owe | eno, | nacoot | seston face | ney | aorsam | totes fcr | Goce |) Eateae | sarees Keowe: | reme | cage, |, Sonne a iets | casnaneReseatch Journal of Blogcal Sconces Inflrecene> Moret 3 an - Mar 2009, pp, 168177 ‘Table 3. Effect of Groundnut Oileake Medium Supplemented! with Different Carbon and Nitrogen Sources, on the Biomass Production and Protein Content of A. aryeae | Gtucose Lactose | Repwone Deurose | Urea Glucose | Uren Lactose | Une Dexwose | Yet exact | 0.4220.03 worse | Yeastestact | 0.382001 Lacsoee | Yea ett eee nthe Sesmum ofleake mediam supplementation seudies only A. nigerresponded well. when niuogen source alone ‘Was supplemented CTuble 4, Fig.2) sndwhen both carbon and nitrogen sources were supplement, the maximum m YBa ima (en), ‘oad 200) | 287 xOOT os02002 | a0as001 oreo | 20320m oasoo | 2082001 c@soa | 2022001 oxsom | 0822001 022003 | 20200 ox+001 | 2034003 osszom | oxseom os2002 | 2082001 osoo | oss2001 0322001 | 0732001 oa20m | Leno 0364003 | 0a3 2001 Dss2001 | 0724001 o4exo02 | 10r+a02 oazs0or | 0722001 es6z001 | os2s001 ass2001 732001 204002 | o61z002 biomass and protein produ mn Were recorded in Dextrose and NaNO, supplemented medium, which were 2.824002 g/l and 9.340.01 ig/ml respectively (Table 5)Meporeden Sra Sela Mar 3003. pp 168-177 ective Producton of Biomans of Apert wp Using Difeent Ottakes “Table 4. Effect of Sezamum Cilcake Medium Supplemented with Different Nitrogen Sources, on the Biomass Production and Proteia Content of A. niger and A. oryzae ‘Oaganemused | Nitogen Source | Total prowin Bay lomass S “gie ——— Nanos Oss 2001 Tos £0.05 (NH, SO. ves 20.02 Vos 20.02 Niel bea 20.03 oe 20.01 KNOs 59 2001 083 20.03 Peprone Oss t0.02 be 20.02 ‘Urea oe 2001 8020.01 Yeateurct | 0562001 bx2 20.01 A onsae NaNO 8.69 5003 tio 2002 eatin. So. 053 Fo02 Yo2 20.01 NELACI 0.58 50.03 og 2001 KNOs Bae 2001 Os S001 : Peprone ots £0.02 Bio 20.01 Brea 054 2001 Oat 2001 fs eat extrmce | 0502003 3003 Figure 2. fect of Sesamum Oilcake Medium Supplemented with Different Nitrogen Sources, ‘on the Biomass Production aad Protein Content of A. niger and A. oryane | i 08 sce nediy {orescence Pore rien Mar 3008 pp. (68-177 "Table 5. Effect of Sesamum Oilcake Medium Supplemented with Different Carbon ard ‘Nitrogen Sources, on the Biomass Production and Protein Content of A. niger ‘Calton Source | Nitrogen Soaice | Total provein Dry Biomass zim) (ohm) Dexirore NaNGS | 993 007 Tee 0m gh (NH), SO, 0.83 20.01 1820.01 gf NECI 0.76 #0.02 1.08 20.01 KNO; 073 20.03 1022001 Peptone 0.68 20.01 a £0.02 Urea os42001 0-72 #001 ‘Yeast extract 052 20.02 om 20.03 Glucose NaNoy 0.92 20.01 2.63 20.02 (NH; SO, 0.85 20.02 1722001 NLC. pes 20.02 702002 KNO, 0.76 20.01 x2 4002 Peptone 0.71 20.01 1.512002 Urea 0.683 0.01 1.01 £0.03 Yeastextract 058 20.01 1.00 +002 Lactose Nanos 0.902 0.01 2532001 | (NH02 80, 0.80 2.0.01 1524001 NEC 0.76 0.03 12 £003 KNO, 0.62 20.02 1.322002 0.60 = 0.02 1ats001 0.58 0.01 1302001 0.52 20.01 1.102003 1m the coconut oilcake supplementation studies also maximambiomass and protein productions were reconled A. niger alone responded well, when nitrogen source in dextrose and NaNo, supplemented medium, which lone was supplemented (Table 6 Fig. 3) and when both were found 10 be 2.52:20.02im and 9220.0 igienl carbon and nitrogen sources were supplemented the respectively (Table 7).Reseach Journal of Biological Sciences Tnvloescence 1 Fort 3. an” Mar 2009, 9p. 168-177 nective Production of Biomass of Aspergillus sp Using Diferent Oileakes ‘Table 6. Effect of Coconut Oileake Medium Supplemented with Different Nitrogen Sources, on the Biomass Production and Protein Content of A. niger and A. oryzae ‘Organiam wed] Nitrogen Source | Total provein | Day biomass] agi) | _ (w/e) a niger NaNO 0832001 | 1a3=007 (NH)2S0, | 0.632002 | 1322001 | NHACI 0.532003 | 083+ 0.02 KNO: 0.522003 | 0922003 Peptone 0.712002 | 0832001 Ures 0633002 | 0732005 Yeastextract | 0.612001 | O81 +002 A. onvvae ‘NaNOs 0.81 2001 108 £0.01 (WH);S0, | 0.732003 103 50.01 | sites 0.63 #001 083 +001 KNOs 0612002 | 0922002 | peptone 0.532002 | 0832002 Urea 043003 | 063+001 Yeastextract_| 0.58003 | _04220.02 Figure 3. Effect of Coconut Oilcake Medium Supplemented with Different Nitrogen Sources, fon the Biomass Production and Protein Content of A. niger and A. onzae 06 ~ | o4 | a —retairrtein | ° poems | SEPSEERSSES RES | | gg2f2°532253°% Z a2 i |oscene Hove in“ ioe S00,pp 168t77 “Table 7. Eifect of Coconst Oilcake Mealum Supplemented with Different Carbon and Nitogen ‘Sources, en the Biomass Production and Protein Content of A. niger ‘Yeast extract “The present study reveals that among the 180 species of Aspergillus studied A. niger bas more capacity to prodace igh biomass and prein comtent in groundnut oilcake medium supplemented with NaNO, as nitrogen seurce and Glucose as earhon source (Qlinkishide et, 2008) Oconee fatty nctinprotet ‘Carbon Source | Niuogen Source | Total Proveia | Dey Biomass Dextose NaNO; | oste0or | 252=002 exny:s0, | ososocr | 2432002 ILC omsoa | 182-002 KNO; osiz002 | ost soo Peptone o7s2002 | 080001 Urea oes003 | 071200. Yeatextme: | o7is001 | o«2soor | Glucose NaNOs osz2o01 | 2072001 | @m).so, | 072003 | 2002001 NILCT oerso02 | 132001 NO} omssoo1 | 1432002 Peptone omsoor | os1z0cz Urea «22002 | os2+001 Yeas ext | goz001 | 0712001 Lactose NaNO oet2001 | o8is001 exiip; 50, | 0602001 | o712001 | NIL oss2001 | 0612001 | KNO, oaszoo1 | osss001 Peptone oaszoo | 0422001 Urea cazzoo | oszsoo1 o.1 001 432001 and taditinally used 3 fer fo farm animals (Sing et ‘a, 2003), They ate ss ase in the area of fermentation Forenzyme prouction. They are cheaper than Iborstory chemicals which arc uscd forthe preparation of mck Hence they can be recommended as cost effective prodiction mes for high Biome and protein NarvestReseach Jour of Biopial Scenes Tnflaescenc I, le Jan Mar 208 pp 168-177 tect Fraction of Baas Aspergla p Usig Dire: Oakes References Abbott E V (1992). The occurence and action of fungi in soll, Sot Sci, 16, 207-216. 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