Production of limonoate A-ring lactone by immobilized limonin D-ring lactone hydrolase

Abstract

Limonin D-ring lactone hydrolase, the enzyme catalysing the reversible lactonization/hydrolysis of D-ring in limonin, has been purified from Citrus seeds and immobilized on Q-Sepharose to produce homogeneous limonoate A-ring lactone solutions. The immobilized limonin D-ring lactone hydrolase showed a good operational stability and was stable after sixty-seventy operations and storing at 4°C for six months.

Isolation and characterization of limonoate and nomilinoate A-ring lactones

“…The existence of 3 and 4 has been known for decades, however their inherent instability has precluded their isolation and characterization as pure compounds Maier and Margileth, 1969;Merino et al, 1996Merino et al, , 1997. We now report the first isolation and characterization of 3 and 4.…”

Purification and characterization of limonoate dehydrogenase from Rhodococcus fascians

“…To circumvent that problem we have used immobilized LDRL hydrolase. This approach has allowed us to obtain practically homogeneous LARL solutions, with limonin contaminations lower than 2% (21). In any case, the presence of limonin does not affect the LDase assay, since the enzyme is inactive towards that limonoid due to its closed D ring (2).…”

“…For production of LARL, purified limonoate D-ring lactone (LDRL) hydrolase immobilized on a Q-Sepharose column (6.5 by 1.6 cm) was used (21). The column was equilibrated with 100 mM Tris-HCl, pH 8.0.…”

Crystallization and preliminary X-ray diffraction study of lactonohydrolase from Fusarium oxysporum

“…3.1.1.46), and actinomycin lactonase (E.C. 3.1.1.39) are involved in the degradation pathways for vitamin B 6 (Jong & Snell, 1986), aromatic compounds (Kosuge & Conn, 1962;Casellas et al, 1997), limonoids (Hasegawa et al, 1980;Merino et al, 1996) and actinomycin (Hou & Perlman, 1970), respectively. As regards molecular biochemical studies, only the gene encoding gluconolactonase from Zymomonas mobilis has been cloned and sequenced (Kanagasundaram & Scopes, 1992), and there have been no reports of the three-dimensional structure of any lactonohydrolase.…”