Absorption Spectroscopic Studies of Prodigiosin Extracted from Serratia Marcescens Strain

Serratia marcescens 균주로부터 추출한 Prodigiosin의 흡수분광학적 연구

Abstract

The red pigment extracted from Serratia marcescens 2354 (ATCC 25419) was prodigiosin (PG), which was dissolved in methanol and measured for ultraviolet and visible light absorption spectra. It was the typical absorption spectrum of PG in an acid solution with ${\lambda}_{max}=537nm$. When the concentration of PG was increased from $1.0{\times}10-5M$ to $9.0{\times}10-5M$ in the methanol solution, the absorption intensity at 537 nm was increased, the absorption intensity at 467 nm was decreased, and the isosbestic point at 500 nm was observed. This phenomenon can be regarded as a result of reversible acid-base equilibrium reaction considering 537 nm and 467 nm of PG absorption band in acid and base solution respectively and isosbestic point of 500 nm. On the other hand, when the concentration of PG was reduced from $6.0{\times}10-4$ to $1.0{\times}10-4M$ in acetic acid buffer solution at pH 4.75, a new absorption band with ${\lambda}$ max at 500 nm appeared. This absorption band appears only in the aqueous solution of pH 4.75 and does not appear in the pure methanol solution of the same pH. This is due to the conversion of the PG molecule from the ${\alpha}$-isomer to the ${\beta}$-isomer by $H_2O$. In other words, it was confirmed that the color change of the PG can be caused by the concentration of the solution and the characteristics of the solvent.

Serratia marcescens 2354(ATCC 25419) 균주로부터 추출한 붉은 색소는 prodigiosin (PG)이었고, 이를 methanol에 녹여 자외선 및 가시광선 흡수 스펙트라를 측정한 결과 537 nm의 최대흡수파장 (${\lambda}_{max}$)을 갖는 산성용액에서의 전형적인 PG의 흡수 스펙트라이었다. 또한 methanol 용액에서 PG의 농도를 $1.0{\times}10-5M$에서 $9.0{\times}10-5M$로 증가시키면, 537 nm의 흡수강도는 증가하고 467 nm의 흡수강도는 감소하였으며, 500 nm에서 isosbestic point가 관측되었다. 이러한 현상은 537 nm와 467 nm가 각각 산과 염기용액에서의 PG 흡수대이고, 500 nm의 isosbestic point 등을 고려하면 가역적 산-염기 평형반응에 의한 결과라고 볼 수 있다. 한편 pH, 4.75의 acetic acid 완충용액에서 PG의 농도를 $6.0{\times}10^{-4}M$에서 $1.0{\times}10^{-4}M$로 감소시키면 500 nm에서 ${\lambda}_{max}$를 가지는 새로운 흡수대가 나타난다. 이 흡수대는 pH 4.75의 수용액에서만 나타나는 것으로 같은 pH의 순수한 methanol 용액에서는 나타나지 않는다. 이는 PG 분자가 $H_2O$에 의해 ${\alpha}$-이성질체에서 ${\beta}$-이성질체로의 전환에 기인하는 것이다. 즉 PG의 색변화는 용액의 농도 및 용매의 특성에 의해서도 일어날 수 있음을 확인하였다.

Fig. 1. Absorption spectra of PG extracted from Sm. strain at 25 ℃ in methanol.

Fig. 2. The absorption spectra at various concentration of prodigiosin in methanol solution at 25 ℃.

Fig. 3. The absorption spectra at various concentrations of PG in acetic acid buffer solution at 25℃.

Fig. 4. Geometric isomers of prodigiosin.