Determination of molecular mass of the farnesol dehydrogenase from <i>P</i>. <i>minus</i>.

<p>(A) Estimation of native molecular mass of farnesol dehydrogenase by TSK-gel GS3000SW column. Experimental conditions are described in “Materials and Methods”. Standard protein marker (■): thyroglobulin (670 kDa), γ-globulin (158 kDa), ovalbumin (44 kDa), myoglobin (17 kDa), and vitamin B₁₂ (1350 Da). Farnesol dehydrogenase (●). (B) SDS-PAGE analysis of purified farnesol dehydrogenase. Purified enzyme and standard proteins were subjected to electrophoresis in the presence of SDS with a 12.5% polyacrylamide gel. The PageRuler™ Prestained Protein Ladder, ~10–170 kDa (SM0671) (Fermentas) was used as the molecular marker.</p

List of substrates tested for substrate specificity of <i>P</i>. <i>minus</i> farnesol dehydrogenase.

<p>List of substrates tested for substrate specificity of <i>P</i>. <i>minus</i> farnesol dehydrogenase.</p

Native-PAGE of the purified farnesol dehydrogenase from <i>P</i>. <i>minus</i>.

<p>Purified enzyme (33 μg) was subjected to electrophoresis in the absence of SDS with 12.5% gel at pH 8.8. Protein gel were stained by silver stain (A) and activity stain (B).</p

Summary of purification of farnesol dehydrogenase from <i>P</i>. <i>minus</i> leaves.

<p>Summary of purification of farnesol dehydrogenase from <i>P</i>. <i>minus</i> leaves.</p

Substrate specificity, coenzyme specificity, and kinetic parameters of <i>P</i>. <i>minus</i> farnesol dehydrogenase.

<p>n.d- not determined.</p><p>Substrate specificity, coenzyme specificity, and kinetic parameters of <i>P</i>. <i>minus</i> farnesol dehydrogenase.</p

Identification of tryptic peptides from <i>P</i>. <i>minus</i> farnesol dehydrogenase.

<p>¹ (XP_002534918.1) putative 3-oxoacyl-[acyl-carrier-proten] reductase of <i>Ricinus communis</i></p><p>² (XP_009144454.1) predicted carbonyl reductase family member 4 of <i>Brassica rapa</i></p><p>³ (XP_009415001.1) predicted uncharacterized oxidoreductase YMR226C isoform X1 of <i>Musa acuminata subsp</i>. <i>Malaccensis</i></p><p>⁴ (XP_009792301.1) Predicted dehydrogenase/reductase SDR family member 7-line of <i>Nicotiana sylvestris</i></p><p>⁵ (AAB62297.1) <i>p</i>-cumic alcohol dehydrogenase of <i>Pseudomonas putida</i></p><p>⁶ (ADB03640.1) NADP⁺-dependent farnesol dehydrogenase 2 of <i>Aedes aegypti</i></p><p>⁷ (AEE86213.1) Rossmann-fold NAD(P)-binding domain-containing protein of <i>Arabidopsis thaliana</i></p><p>⁸ (AAU20370.1) (-)-isopiperitenol dehydrogenase of <i>Mentha x piperita</i></p><p>⁹ (XP_005650109.1) NAD(P)-binding protein from <i>Coccomyxa subellipsoidea</i> C-169</p><p>¹⁰ (XP_004231947.1) predicted NADPH-dependent -1-acylihydroxyacetone phosphate reductase-like of <i>Solanum lycopersicum</i></p><p>¹¹ (ABA99950.2) oxidoreductase, short chain dehydrogenase/reductase family protein, expressed from <i>Oryza sativa Japonica</i> group</p><p>¹² (CCF55024.1) geraniol dehydrogenase of <i>Castellaniella defragrans</i></p><p>¹³ (KDO39154.1) hypothetical protein CISIN_1g030018mg of <i>Citrus sinensis</i></p><p>¹⁴ (XP_011091715.1) predicted (+)-neomenthol dehydrogenase of <i>Sesamun indicum</i></p><p>¹⁵ (AII08455.1) geraniol dehydrogenase <i>(Rhodococcus opacus)</i></p><p>¹⁶ (AHZ20737.1) farnesol dehydrogenase, partial <i>(Diploptera punctata</i>)</p><p>^a nd.–not determined</p><p>Identification of tryptic peptides from <i>P</i>. <i>minus</i> farnesol dehydrogenase.</p

Effects of temperature and pH.

<p>(A) Effects of temperature on enzyme activities of farnesol dehydrogenase and stability of the enzyme. The temperature stability was determined by incubating the purified enzymes at a temperature in the range of 25–70°C for 10 min at pH 7.5 (100 mM tricine-NaOH containing 2.5 mM 2-ME). The residual farnesol dehydrogenase activity was assayed as described in “Materials and Method”. The optimal temperature was determined by performing the standard enzyme assay as described in “Materials and Methods,” except that the reaction temperature was varied. Thermo stability (●), optimal temperature (■). (B) Effect of pH on enzyme activity of farnesol dehydrogenase. Enzyme activity was assayed under the standard assay conditions, except that the following buffers were used at a final concentration of 100 mM in the incubation mixture: citrate buffers (■), potassium phosphate buffers (×), Tris-HCl buffers (○), glycine-NaOH buffers (▲), and carbonate buffers (●).</p