: Chitinous materials from marine byproducts elicit great interest among biotechnologists for
their potential biomedical or agricultural applications. In this study, four kinds of marine chitinous
materials (squid pens, shrimp heads, demineralized shrimp shells, and demineralized crab shells)
were used to screen the best source for producing chitosanase by Paenibacillus macerans TKU029.
Among them, the chitosanase activity was found to be highest in the culture using the medium
containing squid pens as the sole carbon/nitrogen (C/N) source. A chitosanase which showed
molecular weights at 63 kDa was isolated from P. macerans cultured on a squid pens medium.
The purified TKU029 chitosanase exhibited optimum activity at 60 ◦C and pH 7, and was stable at
temperatures under 50 ◦C and pH 3-8. An analysis by MALDI-TOF MS revealed that the chitosan
oligosaccharides (COS) obtained from the hydrolysis of water-soluble chitosan by TKU029 crude
enzyme showed various degrees of polymerization (DP), varying from 3–6. The obtained COS
enhanced the growth of four lactic acid bacteria strains but exhibited no effect on the growth of E. coli.
By specialized growth enhancing effects, the COS produced from hydrolyzing water soluble chitosan
with TKU029 chitinolytic enzymes could have potential for use in medicine or nutraceuticals.