In this study, we examined how land use and urbanization changes in adjacent areas affected biological productivity and carbon cycling in a lake ecosystem over 100 years and how these changes are reflected in carbon isotope variations. We performed radiocarbon (14C) activity and stable carbon isotope ratio analysis in two organic fractions: humin and humic acids of lake sediment. Additionally, we performed pigment and diatom analysis and determined the carbonate and organic matter (OM) content in sediments. Over the last century, the estimated 14C reservoir age in both sediment organic fractions varied from 1136 ± 112 yr to 5733 ± 122 yr. The increase in the reservoir age by 1175 ± 111 yr was related with higher inputs of pre-aged organic carbon and 14C depleted hard water due to the opening of the channel connecting two lakes. Nuclear weapons tests caused an increase in the reservoir age of up to 5421 ± 135 yr and 5733 ± 122 yr in humin and humic acids, respectively. 13C values in the humic acid fraction showed a tendency to decrease, depending on the content of autochthonous versus allochthonous OM in sediments, while changes in the sources of OM had a minor impact on the stable carbon isotope composition in the humin fraction.