A comparison of non-hydrolytic methods for extracting amino acids and proteins from coastal marine sediments

Advances in analytical techniques now allow for the potential analysis of intact peptides and proteins isolated from marine sediments. However, there is no established technique for the extraction of macromolecular materials from marine sediments. Six different methods for extracting the amino acid component from coastal marine sediments were compared to the standard hot acid hydrolysis technique for their percent recovery and amino acid composition. The standard hot acid hydrolysis on dried, whole sediments released the greatest concentration of total amino acids (PS-THAA; 3.52 mg gdwt^− 1 ± 10% (SMD)), yet this only accounted for 22% of the total nitrogen in Puget Sound sediments (Washington, USA). Repeated hydrolysis of the same samples did not improve the recovery of nitrogen by more than an additional 10%. Base extraction (0.5 N NaOH) was the second best method for recovering amino acid nitrogen, releasing 60% of the Puget Sound total hydrolyzable amino acids (PS-THAA) (corresponding to 13% of the total sedimentary nitrogen), and has the advantage that it does not rely on peptide hydrolysis to free the nitrogenous component from the sediment matrix. The amino acid distribution of the 0.5 N NaOH extract was not significantly different than the initial THAA. Other non-hydrolyzing methods released lower yields of amino acids (Triton X-100 ≥ hot water > 50 mM NH₄HCO₃ > HF), but might prove to be of use to investigators interested in specific fractions of sedimentary organic nitrogen because these four methods had distinctly different amino acid compositions (enrichments in basic amino acids and depletions in acidic amino acids). Treatments with HF both before and after traditional hydrolysis and/or extractions with base did not release any more of the sedimentary nitrogen. Our results are consistent with the hypothesis that a large fraction of the sedimentary nitrogen (TN) is protected within an organic matrix.     

胶州湾沉积物中氨基酸对有机质降解及细菌源贡献的指示作用解析

基于对胶州湾表层沉积物中总可水解氨基酸（THAA）的含量、组成、构型及分布特征的系统研究,通过氨基酸碳氮归一化产率（THAA-C%,THAA-N%）、降解因子DI、反应活性指数RI以及D型氨基酸占比（D-AA%,摩尔百分比）等指标结合碳氮比（TOC/TN）、碳稳定同位素（δ13C）探析了胶州湾沉积物中有机质的来源与降解状态,利用细菌源有机质及胞外肽酶活性（EEA）探讨了微生物在有机质迁移转化过程中的作用与贡献。结果表明,胶州湾表层沉积物中氨基酸平均含量为（7.60±3.64）μmol/g,在陆源与海源混合影响下,其水平分布呈现湾内高于湾外、湾内东部高于西部的特点,表明湾内东部陆源输入对沉积物THAA具有较高贡献。THAA-C%、THAA-N%、DI、RI以及D-AA%等指示因子均显示胶州湾表层沉积物中有机质的降解程度呈现湾外高于湾内、湾内东部高于西部的变化趋势,有机质来源、微生物活性与上覆水水深共同影响了有机质的降解程度。胶州湾表层沉积物中细菌源有机碳的贡献率为（29.35±18.73）%,其水平分布显示出湾内西部与湾外相近且高于湾内东部的特点。细菌胞外肽酶活性（EEA）平均为（0.81±1.31）nmol/（g·h）（以MCA计）,整体分布趋势与细菌贡献率相反,呈现湾内东部高于湾内西部和湾外的特性。沉积物中有机质的不同海源、陆源占比决定了有机质的可降解性,而有机质的降解程度进一步影响了细菌源有机质的贡献与胞外肽酶活性。     

Seasonal variations in the production and consumption of amino acids by coastal microplankton

A seasonal study on coastal microplankton was conducted in surface waters near Boothbay Harbor, Maine. Phytoplankton biomass, particulate production and extracellular organic release were examined in conjunction with microheterotrophic biomass and the uptake and respiration of amino acids. In situ dissolved free amino acid (DFAA) concentrations were also determined. Several phytoplankton blooms occurred throughout the year, in mid-summer, late autumn and in mid-winter. Heterotrophic activity and biomass paralleled phytoplankton extracellular release more closely than either phytoplankton particulate production or biomass. DFAA concentrations were not wholly dependent on extracellular release. Heterotrophic uptake did not appear to be dependent on DFAA concentrations but rather on rates of production of DOC by phytoplankton.