Isolation of Erwinia chrysanthemi mutants altered in pectinolytic enzyme production

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Abstract
Summary: Various mutations in the pectin catabolic pathway of Erwinia chrysanthemi were isolated by selection of Mu‐lac insertions, resulting in expression of the lac genes inducible by pectin degradation products. This approach allowed us to isolate lacZ fusions with the genes pelC, pelD, ogl and pem, encoding pectate lyases PLc and PLd, oligogalacturonate lyase and pectin methytesterase, respectively. Moreover, we obtained mutations affecting the regulation of pectinolytic enzymes; a locus called peel appeared to be involved in induction of pectate tyases and pectin methylesterase. A second locus, called pect, may encode an activator protein acting on pectate lyase production. Both peel and pecL expression are induced in the presence of pectic polymers. The expression of the pem gene was studied in more detail by analysis of the pem‐lacZ fusions. The expression of pem appears to be controlled by the negative regulatory gene kdgR, which controls alt the genes involved in pectin degradation (pem, pel, ogl, kduD, kduf, kdgK, kdgA). This study confirmed that 2‐keto‐3‐deoxy‐gluconate is a key intermediate for the induction of the pectin catabolic pathway. The three genes pem, pelD and pecl were localized in the same region, near the ade‐377 marker on the genetic map of the E. chrysanthemi strain 3937. The pem gene was located more precisely on an 18kb DNA fragment containing the pelADE cluster. However, this 18 kb DNA fragment did not complement the pecl mutation. The pecL mutations were located near the ile‐2 marker on the genetic map of E. chrysanthemi strain 3937.