Cathepsin L (CPL) cysteine protease is a proteolytic enzyme that involves in many biological processes in a wide range of organisms. In free-living nematode Caenorhabditis elegans, CPL plays important roles in embryogenesis and development processes. The CPL protein is also believed to have a role in degradation of blood meal in the gut of parasitic nematodes. Considering this enzyme might play the same functions in parasitic nematodes, CPL became a potential candidate for vaccination against Haemonchus contortus, a gastrointestinal nematode of small ruminants. H. contortus has been shown to have variations in term of morphology and genetic materials that correlated with different hosts and geographical areas. These variations could hinder the development of effective vaccines. Thus, the present study was conducted to clone and characterize recombinant Hc-CPL-1 from H. contortus isolated from a goat population in Penang, Malaysia. Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify target complimentary DNA (cDNA) from total RNA and protein expression using Escherichia coli expression system was performed from constructed cDNA clone library. The identity of each protein band was confirmed by liquid chromatography-mass spectrometry (LC-MS) analysis followed by De novo sequencing and database matching. The protein structure and its evolutionary relationship were also studied using several bioinformatics approaches. Basic local alignment search tool (BLAST) analysis of the strain retrieved from clone library showed 99% sequence similarity to the Haemonchus cathepsin L cysteine protease and a 47 kDA protein was successfully expressed. Bioinformatic analysis indicated that this protease has a close relationship with Dv-CPL-1, Sv-CPL-1 and Ce-CPL-1. These data might provide an insight on manipulating this enzyme for future novel vaccine development.