Multidrug resistance, especially multidrug efflux mechanisms that extrude structurally unrelated cytotoxic compounds from the cell by multidrug transporters, is a serious problem and one of the main reasons for the failure of therapeutic treatment of infections by pathogenic microorganisms as well as of cancer cells. Streptococcus mutans is considered one of the primary causative agents of dental caries and periodontal disease, which comprise the most common oral diseases. A fragment of chromosomal DNA from S. mutans KCTC3065 was cloned using Escherichia coli KAM32 as host cells lacking major multidrug efflux pumps. Although E. coli KAM32 cells were very sensitive to many antimicrobial agents, the transformed cells harboring a recombinant plasmid became resistant to several structurally unrelated antimicrobial agents such as tetracycline, kanamycin, rhodamin 6G, ampicillin, acriflavine, ethidium bromide, and tetraphenylphosphonium chloride. This suggested that the cloned DNA fragment carries a gene encoding a multidrug efflux pump. Among 49 of the multidrug-resistant transformants, we report the functional gene cloning and characterization of the function of one multidrug efflux pump, namely MdeA from S. mutans, which was expressed in E. coli KAM32. Judging from the structural and biochemical properties, we concluded that MdeA is the first cloned and characterized multidrug efflux pump using the proton motive force as the energy for efflux drugs.

To use oil palm empty fruit bunch (EFB) for cellulase production, a novel fungus was isolated from moistened EFB. This fungus was classified as Penicillium sp. by sequence analysis of the internal transcribing space and it was named Penicillium sp. GDX01. The Penicillium sp. strain secreted cellulases under solid-state fermentation of EFB. The fermentation conditions were optimized for maximal enzyme production. Of the different substrates tested, both EFB and rice straw gave the maximum production of filter paper activity (FPase). Five percent yeast extract and 40-50% initial moisture content were found to be optimal for enzyme production. In addition, the pretreatment of EFB with NaOH before fermentation inhibited the cellulase production of Penicillium sp. GDX01. Saccharification of pretreated EFB by cellulases from Penicillium sp. GDX01 resulted in a more than 80% release of glucose during a 72 h incubation, which is a better result than when using Celluclast 1.5L without the addition of β-glucosidase. Our results show that the cellulases produced by Penicillium sp. GDX01 are more efficient at the saccharification of EFB than Celluclast 1.5 L.