Application Notes

8 APPLICATION NOTE ©2017 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved. Figure 6: Formulation screening of TAKA. In order to find optimal conditions for an increased thermal stability of TAKA, its thermal unfolding was monitored under 16 different additive conditions. A clear shi of T m towards higher values is visible in the fluorescence ratio plots for each additive. Quantification of T m under different conditions renders the addition of 40 % sucrose most effective, while glycerol and trehalose have the weakest effect. 20 30 40 50 60 70 80 90 Sorbitol (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Sucrose (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Glycerol (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Trehalose (% w/v) 0 10 20 30 40 0 10 20 30 40 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 Trehalose Glycerol Sucrose Sorbitol % additive (w/v) A Aspergillus oryzae a-amylase formulation screen B Aspergillus oryzae a-amylase Temp (°C) Temp (°C) Temp (°C) Temp (°C) T (°C) m F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 Aspergillus oryzae α-amylase B % additive (w/v) 20 30 40 50 60 70 80 90 Sorbitol (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Sucrose (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Glycerol (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Trehalose (% w/v) 0 10 20 30 40 0 10 20 30 40 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 Trehalose Glycerol Sucrose Sorbitol % additive (w/v) A Aspergillus oryzae a-amylase formulation screen B Aspergillus oryzae a-amylase Temp (°C) Temp (°C) Temp (°C) Temp (°C) T (°C) m F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 A Aspergillus oryzae α-amylase formulation screen Temp (°C) Temp (°C) 20 30 40 50 60 70 80 90 Sorbitol (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Sucrose (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Glycerol (% w/v) 0 10 20 30 40 20 30 40 50 60 70 80 90 Trehalose (% w/v) 0 10 20 30 40 0 10 20 30 40 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 Trehalose Glycerol Sucrose Sorbitol % additive (w/v) A Aspergillus oryzae a-amylase formulation screen B Aspergillus oryzae a-amylase Temp (°C) Temp (°C) Temp (°C) Temp (°C) T (°C) m F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 F350 / F330 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 0.90 0.85 Temp (°C) Temp (°C)

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