Ciri inokulum bagi pengkulturan kulat oleaginus pencilan tempatan, Cunninghamella bainieri 2A1 dibangunkan dengan mengenal pasti kesan jenis, umur dan saiz inokulum terhadap pertumbuhan, penghasilan lipid dan GLA. Pengkulturan dijalankan pada suhu 30ºC dengan kadar goncangan 250 rpm dalam kelalang goncangan 500 mL yang mengandungi 200 mL medium terhad nitrogen. Inokulum spora didapati lebih sesuai berdasarkan produktiviti penghasilan lipid yang tinggi iaitu 0.71 (g/L/hari) berbanding penggunaan inokulum sel vegetatif vegetatif 24 jam dan 48 jam yang masingmasing memberikan produktiviti hanya 0.51 dan 0.45 (g/L/hari). Selain itu, penghasilan GLA (5.3 × 10-2 g/g biojisim tanpa lipid) dalam kultur yang dimulakan dengan inokulum spora (1 × 105 spora/mL) didapati lebih tinggi sebanyak 23% berbanding inokulum sel vegetatif. Kepekatan spora sebanyak 1 × 103 spora/mL menghasilkan morfologi pellet bersaiz 1.04 mm dan berkadaran dengan kandungan lipid dan GLA masing-masing sebanyak 40% (g/g biojisim) dan 8.34 × 10-2 (g/g biojisim tanpa lipid).
The effects of ammonium tartrate and glucose concentration on biomass, lipid and GLA accumulation in Cunninghamella sp. 2A1 were investigated using Response Surface Methodology (RSM). Cultivation was carried out in 250 mL shake flask containing 100 mL of nitrogen limiting medium (with various combinations of concentration of ammonium tartrate (1-3 g/L) and glucose (30-60 g/L) at 30°C and 250 rpm agitation for 120 h. The concentration of both compounds significantly affected the biomass, lipid and GLA yield (p<0.05), with the production of each of them being represented by quadratic models. Higher concentration of ammonium tartrate and glucose (2.99 and 59.33 g/L, respectively) was required for enhanced biomass production whereas low nitrogen content with excess glucose was otherwise favoured for lipid and GLA production. Ammonium tartrate and glucose concentration at 1 and 43 g/L, respectively were estimated by the model and proven to give the highest lipid production and GLA yield of 31.06 % (g/g biomass) and 4.15 ×10-2 (g/g lipid less biomass), respectively